Systems and methods for interior energy-activation from an exterior source

ABSTRACT

A method and a system for producing a change in a medium. The method places in a vicinity of the medium at least one energy modulation agent. The method applies an initiation energy to the medium. The initiation energy interacts with the energy modulation agent to directly or indirectly produce the change in the medium. The system includes an initiation energy source configured to apply an initiation energy to the medium to activate the energy modulation agent.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. Ser. No. 14/635,677 filedMar. 2, 2015. U.S. Ser. No. 14/635,677 is a continuation of U.S. Ser.No. 14/157,039 filed Jan. 16, 2014. U.S. Ser. No. 14/157,039 is acontinuation of U.S. Ser. No. 13/713,974 filed Dec. 13, 2012 which is acontinuation of Ser. No. 12/401,478 filed Mar. 10, 2009, the entirecontents of each are incorporated herein by reference. This applicationis related to provisional Ser. No. 60/910,663, filed Apr. 8, 2007,entitled “METHOD OF TREATING CELL PROLIFERATION DISORDERS,” andnon-provisional Ser. No. 11/935,655, filed Nov. 6, 2007, entitled“METHOD OF TREATING CELL PROLIFERATION DISORDERS,” the contents of eachof which are hereby incorporated herein by reference. This applicationis related to provisional Ser. No. 61/035,559, filed Mar. 11, 2008,entitled “SYSTEMS AND METHODS FOR INTERIOR ENERGY-ACTIVATION FROM ANEXTERIOR SOURCE,” the entire contents of which are hereby incorporatedherein by reference.

THE NAME OF THE PARTIES TO A JOINT RESEARCH AGREEMENT

Immunolight, LLC and Duke University are parties to a joint researchagreement.

BACKGROUND OF THE INVENTION

Field of Invention

The invention relates to methods and systems for generating in theinterior of a medium or body radiant energy for producing a change inthe properties of a medium or body by exposure to the radiation.

Discussion of the Background

Presently, light (i.e., electromagnetic radiation from the radiofrequency through the visible to the x-ray and gamma ray wavelengthrange) activated processing is used in a number of industrial processesranging from photoresist curing, to on-demand ozone production, tosterilization, to the promotion of polymer cross-linking activation(e.g. in adhesive and surface coatings) and others. Today, lightactivated processing is seen in these areas to have distinct advantagesover more conventional approaches. For example, conventionalsterilization by steam autoclaving or in food processing bypasteurization may unsuitably overheat the medium to be sterilized. Assuch, light activated curable coatings are one of the fastest growingsectors in the coatings industry. In recent years, this technology hasmade inroads into a number of market segments like fiber optics, opticaland pressure-sensitive adhesives, and automotive applications like curedtopcoats, and curable powder coatings. The driving force of thisdevelopment is mostly the quest for an increase in productivity of thecoating and curing process, as conventional non light activated adhesiveand surface coatings typically require 1) the elimination of solventsfrom the adhesive and surface coatings to produce a cure and 2) atime/temperature cure which adds delay and costs to the manufacturingprocess.

Moreover, the use of solvent based products in adhesive and surfacecoatings applications is becoming increasingly unattractive because ofrising energy costs and stringent regulation of solvent emissions intothe atmosphere. Optimum energy savings as well as beneficial ecologicalconsiderations are both served by radiation curable adhesive and surfacecoating compositions. Radiation curable polymer cross-linking systemshave been developed to eliminate the need for high oven temperatures andto eliminate the need for expensive solvent recovery systems. In thosesystems, light irradiation initiates free-radical cross-linking in thepresence of common photosensitizers.

However, in the adhesive and surface coating applications and in many ofthe other applications listed above, the light-activated processing islimited due to the penetration depth of light into the processed medium.For example, in water sterilization, ultraviolet light sources arecoupled with agitation and stirring mechanisms in order to ensure thatany bacteria in the water medium will be exposed to the UV light. Inlight-activated adhesive and surface coating processing, the primarylimitation is that the material to be cured must be directly exposed tothe light, both in type (wavelength or spectral distribution) andintensity. In adhesive and surface coating applications, any “shaded”area will require a secondary cure mechanism, increasing cure time overthe non-shaded areas and further delaying cure time due to the existentof a sealed skin through which subsequent curing must proceed (i.e.,referred to as a cocoon effect).

As described in incorporated by reference Ser. No. 11/935,655, it iswell recognized that a major problem associated with the existingmethods of diagnosis and treatment of cell proliferation disorders is indifferentiation of normal cells from target cells. Such targetspecificity is difficult to achieve by way of surgery since the strategythere is simply to cut out a large enough portion of the affected areato include all diseased cells and hope that no diseased cells havespread to other distant locations.

With chemotherapy, while some degree of differentiation can be achieved,healthy cells are generally adversely affected by chemo-agents. As insurgery, the treatment strategy in chemotherapy is also to kill off alarge population of cells, with the understanding that there are farmore normal cells than diseased cells so that the organism can recoverfrom the chemical assault.

Radiation therapy works by irradiating cells with high levels of highenergy radiation such as high energy photon, electron, or proton. Thesehigh energy beams ionize the atoms which make up a DNA chain, which inturn leads to cell death. Unlike surgery, radiation therapy does notrequire placing patients under anesthesia and has the ability to treattumors deep inside the body with minimal invasion of the body. However,the high doses of radiation needed for such therapies damages healthycells just as effectively as it does diseased cells. Thus, similar tosurgery, differentiation between healthy and diseased cells in radiationtherapy is only by way of location. There is no intrinsic means for aradiation beam to differentiate between a healthy cell from a diseasedcell either.

Other methods may be more refined. For example, one form of advancedtreatment for lymphoma known as extracorporeal photopheresis involvesdrawing the patient's blood from his body into an instrument where thewhite cells (buffy coat) are separated from the plasma and the red bloodcells. A small amount of the plasma separated in this process is thenisolated and mixed with a photosensitizer (PS), a drug that can beactivated by light. The buffy coat is then exposed to a light toactivate the drug. The treated blood is then returned to the patient. Inthis example, one may think of the target-specificity problem as beingsolved by separating the blood from the rest of the body where thetarget components are easily exposed.

However, this procedure has its drawbacks; it requires drawing bloodfrom the patient, thus requiring cumbersome machinery to perform and mayrequire blood transfusion in order to maintain the volume of blood flowin the machine. Further, this also limits the size of the patient thatcan be treated, since the extracorporeal volume is great and too muchwithdrawal of blood increases the risk of hypovolemic shock. The methodis also limited to treating blood-born cell proliferation relateddisorders such as lymphoma, and is not capable of treating solid tumorsor other types of non-blood related cell proliferation disorders.

A problem encountered in PDT therapy is the inability to treat targetareas that are more than a few centimeters beneath the surface of theskin without significant invasive techniques, and the fact that PDTtypically operates by generation of sufficient quantities of singletoxygen to cause cell lysis. However, singlet oxygen in sufficientconcentration will lyse not only target cells, but also healthy cellsrather indiscriminately.

Therefore, there still exists a need for better and more effectivetreatments that can more precisely target the diseased cells withoutcausing substantial side-effects or collateral damages to healthytissues, and which are capable of treating even solid tumors or othertypes of non-blood related cell proliferation disorders.

SUMMARY OF THE INVENTION

The invention overcomes the problems and disadvantages of the prior artas described in the various embodiments below.

In one embodiment, there is provided a method and system for producing achange in a medium. The method (1) places in a vicinity of the medium anenergy modulation agent, and (2) applies an initiation energy from anapplied initiation energy source through the artificial container to themedium. The applied initiation energy interacts with the energymodulation agent to directly or indirectly produce the change in themedium. The system further includes an applied initiation energy sourceconfigured to apply the initiation energy to the medium to activate theenergy modulation agent.

As described in incorporated by reference Ser. No. 11/935,655, oneobject of the present invention is to provide a method for the treatmentof a cell proliferation disorder that permits treatment of a subject inany area of the body while being non-invasive and having highselectivity for targeted cells relative to healthy cells.

A further object of the present invention is to provide a method fortreatment of a cell proliferation disorder which can use any suitableenergy source as the initiation energy source to activate theactivatable pharmaceutical agent and thereby cause a predeterminedcellular change to treat cells suffering from a cell proliferationdisorder.

A further object of the present invention is to provide a method fortreatment of a cell proliferation disorder using an energy cascade toactivate an activatable pharmaceutical agent that then treats cellssuffering from a cell proliferation disorder.

A further object of the present invention is to provide a method forgenerating an autovaccine effect in a subject, which can be in vivo thusavoiding the need for ex vivo treatment of subject tissues or cells, orcan be ex vivo.

A further object of the present invention is to provide a computerimplemented system for performing the methods of the present invention.

A still further object of the present invention is to provide a kit anda pharmaceutical composition for use in the present invention methods.

These and other objects of the present invention, which will become moreapparent in conjunction with the following detailed description of thepreferred embodiments, either alone or in combinations thereof, havebeen satisfied by the discovery of a method for treating a cellproliferation disorder in a subject, comprising:

-   -   (1) administering to the subject an activatable pharmaceutical        agent that is capable of effecting a predetermined cellular        change when activated, either alone or in combination with an        energy modulation agent; and    -   (2) applying an initiation energy from an initiation energy        source to the subject,

wherein the applying activates the activatable agent in situ,

thus causing the predetermined cellular change to occur, whereinoccurrence of the predetermined cellular change causes an increase ordecrease in rate of cell proliferation to treat the cell proliferationrelated disorder,

and a kit for performing the method, a pharmaceutical composition, acomputer implemented system for performing the method and a method andsystem for causing an autovaccine effect in a subject.

It is to be understood that both the foregoing general description ofthe invention and the following detailed description are exemplary, butare not restrictive of the invention.

BRIEF DESCRIPTION OF THE FIGURES

A more complete appreciation of the invention and many of the attendantadvantages thereof will be readily obtained as the same becomes betterunderstood by reference to the following detailed description whenconsidered in connection with the accompanying drawings, wherein:

FIG. 1 provides an exemplary electromagnetic spectrum in meters (1 nmequals 10⁻⁹ meters);

FIG. 2 is a table providing a list of photoactivatable agents;

FIG. 3A is a schematic depicting a system according to one embodiment ofthe invention in which an initiation energy source is directed to aself-contained medium for producing changes in the medium;

FIG. 3B is a schematic depicting a system according to anotherembodiment of the invention in which the initiation energy source isdirected to a container enclosing a medium having energy modulationagents disbursed within the medium;

FIG. 3C is a schematic depicting a system according to anotherembodiment of the invention in which the initiation energy source isdirected to a container enclosing a medium having energy modulationagents segregated within the medium;

FIG. 3D is a schematic depicting a system according to anotherembodiment of the invention in which the initiation energy source isdirected to a container enclosing a medium having energy modulationagents segregated within the medium in a fluidized bed configuration;

FIG. 3E and FIG. 3F are graphical representations of the depth ofpenetration of various wavelengths of energy into living tissue.

FIG. 3G illustrates a system according to one exemplary embodiment ofthe present invention.

FIG. 4 illustrates an exemplary computer implemented system according toan embodiment of the present invention.

FIG. 5 illustrates an exemplary computer system (1201) for implementingvarious embodiments of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The invention sets forth a novel method for causing a change in activityof an in a medium that is effective, specific, and able to produce achange to the medium.

Generally, the invention provides methods for producing a change in amedium after generation of radiant light inside the medium. In thismethod, an initiation energy source provides an initiation energy thatpenetrates the medium and induces internal radiation to produce adesired effect in the medium.

In one embodiment, the initiation energy source is applied directly orindirectly to the medium. Within the context of the invention, thephrase “applied indirectly” (or variants of this phrase, such as“applying indirectly”, “indirectly applies”, “indirectly applied”,“indirectly applying”, etc.), when referring to the application of theinitiation energy, means the penetration by the initiation energy intothe medium beneath the surface of the medium and to the activatableagent or energy modulation agents within a medium. In one embodiment,the initiation energy interacts with a previously supplied energymodulation agent which then activates the activatable agent.

Although not intending to be bound by any particular theory or beotherwise limited in any way, the following theoretical discussion ofscientific principles and definitions are provided to help the readergain an understanding and appreciation of the invention.

As used herein, an “activatable agent” is an agent that normally existsin an inactive state in the absence of an activation signal. When theagent is activated by an activation signal under activating conditions,the agent is capable of producing a desired pharmacological, cellular,chemical, electrical, or mechanical effect in a medium (i.e. apredetermined change). For example, when photocatalytic agents areirradiated with visible or UV light, these agents induce polymerizationand “curing” of light sensitive adhesives.

Signals that may be used to activate a corresponding agent may include,but are not limited to, photons of specific wavelengths (e.g. x-rays, orvisible light), electromagnetic energy (e.g. radio or microwave),thermal energy, acoustic energy, or any combination thereof. Activationof the agent may be as simple as delivering the signal to the agent ormay further require a set of activation conditions. For example, anactivatable agent, such as a photosensitizer, may be activated by UV-Aradiation (e.g., by UV-A radiation generated internally in the medium).Once activated, the agent in its active-state may then directly proceedto produce a predetermined change.

Where activation may further require other conditions, mere delivery ofthe activation signal may not be sufficient to bring about thepredetermined change. For example, a photoactive compound that achievesits effect by binding to certain structure in its active state mayrequire physical proximity to the target structure when the activationsignal is delivered. For such activatable agents, delivery of theactivation signal under non-activating conditions will not result in thedesired effect. Some examples of activating conditions may include, butare not limited to, temperature, pH, location, state of the medium, andthe presence or absence of co-factors.

Selection of an activatable agent greatly depends on a number of factorssuch as the desired change, the desired form of activation, as well asthe physical and biochemical constraints that may apply. Exemplaryactivatable agents may include, but are not limited to agents that maybe activated by photonic energy, electromagnetic energy, acousticenergy, chemical or enzymatic reactions, thermal energy, microwaveenergy, or any other suitable activation mechanisms.

When activated, the activatable agent may effect changes that include,but are not limited to an increase in organism activity, a fermentation,a decrease in organism activity, apoptosis, redirection of metabolicpathways, a sterilization of a medium, a cross polymerization and curingof a medium, or a cold pasteurization of a medium.

The mechanisms by which an activatable agent may achieve its desiredeffect are not particularly limited. Such mechanisms may include directaction on a predetermined target as well as indirect actions viaalterations to the biochemical pathways. In one embodiment, theactivatable agent is capable of chemically binding to the organism in amedium. In this embodiment, the activatable agent, is exposed in situ toan activating energy emitted from an energy modulation agent, which, inturn receives energy from an initiation energy source.

Suitable activatable agents include, but are not limited to, photoactiveagents, sono-active agents, thermo-active agents, andradio/microwave-active agents. An activatable agent may be a smallmolecule; a biological molecule such as a protein, a nucleic acid orlipid; a supramolecular assembly; a nanoparticle; or any other molecularentity capable of producing a predetermined activity once activated.

The activatable agent may be derived from a natural or synthetic origin.Any such molecular entity that may be activated by a suitable activationsignal source to effect a predetermined cellular change may beadvantageously employed in the invention.

Suitable photoactive agents include, but are not limited to: psoralensand psoralen derivatives, pyrene cholesteryloleate, acridine, porphyrin,fluorescein, rhodamine, 16-diazorcortisone, ethidium, transition metalcomplexes of bleomycin, transition metal complexes of deglycobleomycin,organoplatinum complexes, alloxazines such as 7,8-dimethyl-10-ribitylisoalloxazine (riboflavin), 7,8,10-trimethylisoalloxazine (lumiflavin),7,8-dimethylalloxazine (lumichrome), isoalloxazine-adenine dinucleotide(flavine adenine dinucleotide [FAD]), alloxazine mononucleotide (alsoknown as flavine mononucleotide [FMN] and riboflavine-5-phosphate),vitamin Ks, vitamin L, their metabolites and precursors, andnapththoquinones, naphthalenes, naphthols and their derivatives havingplanar molecular conformations, porphyrins, dyes such as neutral red,methylene blue, acridine, toluidines, flavine (acriflavinehydrochloride) and phenothiazine derivatives, coumarins, quinolones,quinones, and anthroquinones, aluminum (111) phthalocyaninetetrasulfonate, hematoporphyrin, and phthalocyanine, and compounds whichpreferentially adsorb to nucleic acids with little or no effect onproteins. The term “alloxazine” includes isoalloxazines.

Endogenously-based derivatives include synthetically derived analogs andhomologs of endogenous photoactivated molecules, which may have or lacklower (1 to 5 carbons) alkyl or halogen substitutes of thephotosensitizers from which they are derived, and which preserve thefunction and substantial non-toxicity. Endogenous molecules areinherently non-toxic and may not yield toxic photoproducts afterphotoradiation.

FIG. 1 provides an exemplary electromagnetic spectrum in meters (1 nmequals 1 nanometer). As used herein, an “energy modulation agent” refersto an agent that is capable of receiving an energy input from a sourceand then re-emitting a different energy to a receiving target. Energytransfer among molecules may occur in a number of ways. The form ofenergy may be electronic, thermal, electromagnetic, kinetic, or chemicalin nature. Energy may be transferred from one molecule to another(intermolecular transfer) or from one part of a molecule to another partof the same molecule (intramolecular transfer). For example, amodulation agent may receive electromagnetic energy and re-emit theenergy in the form of thermal energy.

Table 1 in FIG. 2 provides a list of photoactivatable agents that may beused as primary or secondary internal light sources. For example, thephotoactivatable agents could be receptors of X-ray induced emissionsfrom nanoparticles (to be discussed later) and which in turn emit asecondary light. In some mediums, it may be that the excitationwavelengths in Table 1 are transparent to the particular medium and theemission wavelengths are highly absorbent (due to, for example,molecular or solid state band gap transitions). In those cases, thephotoreactive agents in Table 1 would be the primary sources forinternal light generation.

In various embodiments, the energy modulation agent receives higherenergy (e.g. x-ray) and re-emits in lower energy (e.g. UV-A). Somemodulation agents may have a very short energy retention time (on theorder of fs, e.g. fluorescent molecules) whereas others may have a verylong half-life (on the order of minutes to hours, e.g. luminescent orphosphorescent molecules). Suitable energy modulation agents include,but are not limited to, a biocompatible fluorescing metal nanoparticle,fluorescing dye molecule, gold nanoparticle, a water soluble quantum dotencapsulated by polyamidoamine dendrimers, a luciferase, a biocompatiblephosphorescent molecule, a combined electromagnetic energy harvestermolecule, and a lanthanide chelate capable of intense luminescence.Typically, the energy modulation agents induce photoreactive changes inthe medium and are not used for the purpose of exclusively heating themedium.

Various exemplary uses are described in the embodiments below.

The modulation agents may further be coupled to a carrier for targetingpurposes. For example, a biocompatible molecule, such as a fluorescingmetal nanoparticle or fluorescing dye molecule that emits in the UV-Aband, may be selected as the energy modulation agent. The energymodulation agent may be preferably directed to the desired site bysystemic administration into a medium. For example, a UV-A emittingenergy modulation agent may be distributed in the medium by physicalinsertion and or mixing, or by conjugating the UV-A emitting energymodulation agent with a specific carrier, such as a lipid, chitin orchitin-derivative, a chelate or other functionalized carrier that iscapable of concentrating the UV-A emitting source in a specific targetregion of the medium.

Additionally, the energy modulation agent can be used alone or as aseries of two or more energy modulation agents such that the energymodulation agents provide an energy cascade. Thus, the first energymodulation agent in the cascade will absorb the activation energy,convert it to a different energy which is then absorbed by the secondenergy modulation in the cascade, and so forth until the end of thecascade is reached with the final energy modulation agent in the cascadeemitting the energy necessary to activate the activatable agent.Alternatively, one or more energy modulation agents in the cascade mayalso activate additional activatable agents.

Although the activatable agent and the energy modulation agent can bedistinct and separate, it will be understood that the two agents neednot be independent and separate entities. In fact, the two agents may beassociated with each other via a number of different configurations.Where the two agents are independent and separately movable from eachother, they can generally interact with each other via diffusion andchance encounters within a common surrounding medium. Where theactivatable agent and the energy modulation agent are not separate, theymay be combined into one single entity.

The initiation energy source can be any energy source capable ofproviding energy at a level sufficient to activate the activatable agentdirectly, or to provide the energy modulation agent with the inputneeded to emit the activation energy for the activatable agent (indirectactivation). Preferable initiation energy sources include, but are notlimited to, ultraviolet lamps such as UV-A and UV-B lamps, halogenlamps, fiber optic lines, a light needle, an endoscope, self-ballastedmercury vapor lamps, ballasted HID lamps, and any device capable ofgenerating x-ray, y-ray, gamma-ray, or electron beams.

In one embodiment, the initiation energy is capable of penetratingcompletely through the medium. Within the context of the invention, thephrase “capable of penetrating completely through the medium” is used torefer to energy capable of penetrating a container to any distancenecessary to activate the activatable agent within the medium. It is notrequired that the energy applied actually pass completely through themedium, merely that it be capable of doing so in order to permitpenetration to any desired distance to activate the activatable agent.The type of energy source chosen will depend on the medium itself.Exemplary initiation energy sources that are capable of penetratingcompletely through the medium include, but are not limited to, x-rays,gamma rays, electron beams, microwaves and radio waves.

In one embodiment, the source of the initiation energy can be aradiowave emitting nanotube, such as those described by K. Jensen, J.Weldon, H. Garcia, and A. Zettl in the Department of Physics at theUniversity of California at Berkeley (seehttp://socrates.berkeley.edu/˜argon/nanoradio/radio.html, the entirecontents of which are hereby incorporated by reference). These nanotubescan be introduced to the medium, and preferably would be coupled to theactivatable agent or the energy modulation agent, or both, such thatupon application of the initiation energy, the nanotubes would acceptthe initiation energy (preferably radiowaves), then emit radiowaves inclose proximity to the activatable agent, or in close proximity to theenergy modulation agent, to then cause activation of the activatableagent. In such an embodiment, the nanotubes would act essentially as aradiowave focusing or amplification device in close proximity to theactivatable agent or energy modulation agent.

Alternatively, the energy emitting source may be an energy modulationagent that emits energy in a form suitable for absorption by a transferagent or for direct interaction with components of the medium. Forexample, the initiation energy source may be acoustic energy, and oneenergy modulation agent may be capable of receiving acoustic energy andemitting photonic energy (e.g. sonoluminescent molecules) to be receivedby another energy modulation agent that is capable of receiving photonicenergy. Other examples include transfer agents that receive energy atx-ray wavelength and emit energy at UV wavelength, preferably at UV-Awavelength. As noted above, a plurality of such energy modulation agentsmay be used to form a cascade to transfer energy from initiation energysource via a series of energy modulation agents to activate theactivatable agent.

Photoactivatable agents may be stimulated by an energy source throughmechanisms such as irradiation, resonance energy transfer, excitonmigration, electron injection, or chemical reaction, to an activatedenergy state that is capable of producing the predetermined changedesired. One advantage is that wavelengths of emitted radiation may beused to selectively stimulate one or more photoactivatable agents orenergy modulation agents capable of stimulating the one or morephotoactivatable agents. The energy modulation agent is suitablystimulated at a wavelength and energy that causes little or no change tothe medium.

In another embodiment, the photoactivatable agent is stimulated via aresonance energy transfer. Resonance Energy Transfer (RET) is an energytransfer mechanism between two molecules having overlapping emission andabsorption bands. Electromagnetic emitters are capable of converting anarriving wavelength to a longer wavelength. For example, UV-B energyabsorbed by a first molecule may be transferred by a dipole-dipoleinteraction to a UV-A-emitting molecule in close proximity to theUV-B-absorbing molecule. One advantage is that multiple wavelengths ofemitted radiation may be used to selectively stimulate one or morephotoactivatable agents or energy modulation agents capable ofstimulating the one or more photoactivatable agents. With RET, theenergy modulation agent is preferably stimulated at a wavelength andenergy that causes little or no effect to the surrounding medium withthe energy from one or more energy modulation agents being transferred,such as by Foerster Resonance Energy Transfer, to the photoactivatableagents.

Alternatively, a material absorbing a shorter wavelength may be chosento provide RET to a non-emitting molecule that has an overlappingabsorption band with the transferring molecule's emission band.Alternatively, phosphorescence, chemiluminescence, or bioluminescencemay be used to transfer energy to a photoactivatable molecule.

Alternatively, one can apply the initiation energy source to the medium.Within the context of the invention, the applying of the initiationenergy source means the application of an agent, that itself producesthe initiation energy, in a manner that permits the agent to arrive atthe target structure within the medium. In this embodiment, theinitiation energy source includes, but is not limited to, chemicalenergy sources, nanoemitters, nanochips, and other nanomachines thatproduce and emit energy of a desired frequency.

Recent advances in nanotechnology have provided examples of variousdevices that are nanoscale and produce or emit energy, such as theMolecular Switch (or Mol-Switch) work by Dr. Keith Firman of the ECResearch and Development Project, or the work of Cornell et al. (1997)who describe the construction of nanomachines based around ion-channelswitches only 1.5 nm in size, which use ion channels formed in anartificial membrane by two gramicidin molecules: one in the lower layerof the membrane attached to a gold electrode and one in the upper layertethered to biological receptors such as antibodies or nucleotides. Whenthe receptor captures a target molecule or cell, the ion channel isbroken, its conductivity drops, and the biochemical signal is convertedinto an electrical signal. These nanodevices could also be coupled withthe invention to provide targeting of the target cell, to deliver theinitiation energy source directly at the desired site.

In another embodiment, the invention includes the application of theactivatable agent, along with a source of chemical energy such aschemiluminescence, phosphorescence or bioluminescence. The source ofchemical energy can be a chemical reaction between two or morecompounds, or can be induced by activating a chemiluminescent,phosphorescent or bioluminescent compound with an appropriate activationenergy, either outside the medium or inside the medium, with thechemiluminescence, phosphorescence or bioluminescence being allowed toactivate the activatable agent in the medium. The administration of theactivatable agent and the source of chemical energy can be performedsequentially in any order or can be performed simultaneously.

In the case of certain sources of such chemical energy, the applicationof the chemical energy source can be performed after activation outsidethe medium, with the lifetime of the emission of the energy being up toseveral hours for certain types of phosphorescent materials for example.

Yet another example is that nanoparticles or nanoclusters of certainatoms may be introduced such that they are capable of resonance energytransfer over comparatively large distances, such as greater than onenanometer, more preferably greater than five nanometers, even morepreferably at least 10 nanometers. Functionally, resonance energytransfer may have a large enough “Foerster” distance (R₀), such thatnanoparticles in one part of a medium are capable of stimulatingactivation of photoactivatable agents disposed in a distant portion ofthe medium, so long as the distance does not greatly exceed R₀. Forexample, gold nanospheres having a size of 5 atoms of gold have beenshown to have an emission band in the ultraviolet range, recently.

Any of the photoactivatable agents may be exposed to an excitationenergy source provided in the medium. The photoactive agent may bedirected to a receptor site by a carrier having a strong affinity forthe receptor site. Within the context of the invention, a “strongaffinity” is preferably an affinity having an equilibrium dissociationconstant, K_(i), at least in the nanomolar, nM, range or higher. Thecarrier may be a polypeptide and may form a covalent bond with aphotoactive agent, for example. Alternatively, a photoactive agent mayhave a strong affinity for the target molecule in the medium withoutbinding to a carrier.

In one embodiment, a plurality of sources for supplying electromagneticradiation energy or energy transfer is provided by one or more moleculesprovided to the medium. The molecules may emit stimulating radiation inthe correct band of wavelength to stimulate the photoactivatable agents,or the molecules may transfer energy by a resonance energy transfer orother mechanism directly to the photoactivatable agent or indirectly bya cascade effect via other molecular interactions.

In a further embodiment, a biocompatible emitting source, such as afluorescing metal nanoparticle or fluorescing dye molecule, is selectedthat emits in the UV-A band. UV-A and the other UV bands are known to beeffective as germicides.

In one embodiment, the UV-A emitting source is a gold nanoparticlecomprising a cluster of 5 gold atoms, such as a water soluble quantumdot encapsulated by polyamidoamine dendrimers. The gold atom clustersmay be produced through a slow reduction of gold salts (e.g. HAuCl₄ orAuBr₃) or other encapsulating amines, for example. One advantage of sucha gold nanoparticle is the increased Foerster distance (i.e. R₀), whichmay be greater than 100 angstroms. The equation for determining theFoerster distance is substantially different from that for molecularfluorescence, which is limited to use at distances less than 100angstroms. It is believed that the gold nanoparticles are governed bynanoparticle surface to dipole equations with a 1/R⁴ distance dependencerather than a 1/R⁶ distance dependence. For example, this permitscytoplasmic to nuclear energy transfer between metal nanoparticles and aphotoactivatable molecule.

In another embodiment, a UV or light-emitting luciferase is selected asthe emitting source for exciting a photoactivatable agent. A luciferasemay be combined with molecules, which may then be oxygenated withadditional molecules to stimulate light emission at a desiredwavelength. Alternatively, a phosphorescent emitting source may be used.Phosphorescent materials may have longer relaxation times thanfluorescent materials, because relaxation of a triplet state is subjectto forbidden energy state transitions, storing the energy in the excitedtriplet state with only a limited number of quantum mechanical energytransfer processes available for returning to the lower energy state.Energy emission is delayed or prolonged from a fraction of a second toseveral hours. Otherwise, the energy emitted during phosphorescentrelaxation is not otherwise different than fluorescence, and the rangeof wavelengths may be selected by choosing a particular phosphor.

In another embodiment, a combined electromagnetic energy harvestermolecule is designed, such as the combined light harvester disclosed inJ. Am. Chem. Soc. 2005, 127, 9760-9768, the entire contents of which arehereby incorporated by reference. By combining a group of fluorescentmolecules in a molecular structure, a resonance energy transfer cascademay be used to harvest a wide band of electromagnetic radiationresulting in emission of a narrow band of fluorescent energy. By pairinga combined energy harvester with a photoactivatable molecule, a furtherenergy resonance transfer excites the photoactivatable molecule, whenthe photoactivatable molecule is nearby stimulated combined energyharvester molecules. Another example of a harvester molecule isdisclosed in FIG. 4 of “Singlet-Singlet and Triplet-Triplet EnergyTransfer in Bichromophoric Cyclic Peptides,” M. S. Thesis by M. O.Guler, Worcester Polytechnic Institute, May 18, 2002, which isincorporated herein by reference.

In another embodiment, a Stokes shift of an emitting source or a seriesof emitting sources arranged in a cascade is selected to convert ashorter wavelength energy, such as X-rays, to a longer wavelengthfluorescence emission such a optical or UV-A, which is used to stimulatea photoactivatable molecule in the medium.

In an additional embodiment, the photoactivatable agent can be aphotocaged complex having an active agent (which can be a cytotoxicagent if cytotoxicity is needed, or can be an activatable agent)contained within a photocage. In various embodiments, where the activeagent is a cyotoxic agent, the photocage molecule releases the cytotoxicagent into the medium where it can attack non-beneficial “target”species in the medium. The active agent can be bulked up with othermolecules that prevent it from binding to specific targets, thus maskingits activity. When the photocage complex is photoactivated, the bulkfalls off, exposing the active agent. In such a photocage complex, thephotocage molecules can be photoactive (i.e. when photoactivated, theyare caused to dissociate from the photocage complex, thus exposing theactive agent within), or the active agent can be the photoactivatableagent (which when photoactivated causes the photocage to fall off), orboth the photocage and the active agent are photoactivated, with thesame or different wavelengths. Suitable photocages include thosedisclosed by Young and Deiters in “Photochemical Control of BiologicalProcesses”, Org. Biomol. Chem., 5, pp. 999-1005 (2007) and“Photochemical Hammerhead Ribozyme Activation”, Bioorganic & MedicinalChemistry Letters, 16(10), pp. 2658-2661 (2006), the contents of whichare hereby incorporated by reference.

Work has shown that the amount of singlet oxygen necessary to cause celllysis, and thus cell death, is 0.32 H 10⁻³ mol/liter or more, or 10⁹singlet oxygen molecules/cell or more. In one embodiment of theinvention, the level of singlet oxygen production caused by theinitiation energy or the activatable agent upon activation is sufficientto cause a change in a medium, wherein the medium becomes free from anymicroorganisms. Microorganisms include but are not limited to bacteria,viruses, yeasts or fungi. To this end, singlet oxygen in sufficientamounts as described above can be used to sterilize the medium.

For example, medical bottle caps need to be sterilized between the basecap material and the glued seal material which contacts the base of themedical bottle. Because steam autoclaves are insufficient for thispurpose, one embodiment of the invention uses UV luminescing particlesincluded in the adhesive layer when the seal material is applied to thebottle cap. Then, X-ray irradiation becomes capable of curing theadhesive and producing within the adhesive medium UV radiation fordirect sterilization or the production of singlet oxygen or ozone forbiological germicide.

The activatable agent and derivatives thereof as well as the energymodulation agent, can be incorporated into compositions suitable fordelivery to particular mediums. The composition can also include atleast one additive having a complementary effect upon the medium, suchas a lubricant or a sealant.

The carrier can be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), and suitablemixtures thereof. The proper fluidity can be maintained, for example, bythe use of a coating such as lecithin, by the maintenance of therequired particle size in the case of dispersion and by the use ofsurfactants.

Referring to FIG. 3A, an exemplary system according to one embodiment ofthe invention may have an initiation energy source 1 directed at medium4. Activatable agents 2 and an energy modulation agents 3 are dispersedthroughout the medium 4. The initiation energy source 1 may additionallybe connected via a network 8 to a computer system 5 capable of directingthe delivery of the initiation energy. In various embodiments, theenergy modulation agents 3 are encapsulated energy modulation agents 6,depicted in FIG. 3A as silica encased energy modulation agents. As shownin FIG. 3A, initiation energy 7 in the form of radiation from theinitiation energy source 1 permeated throughout the medium 4. A morethorough discussion of the computer system 5 is provided below inreference to FIG. 4. As discussed below in more detail, the initiationenergy source 1 can be an external energy source or an energy sourcelocated at least partially in the medium 4.

In various embodiments, the initiation energy source 1 may be a linearaccelerator equipped with image guided computer-control capability todeliver a precisely calibrated beam of radiation to a pre-selectedcoordinate. One example of such linear accelerators is the SmartBeam™IMRT (intensity modulated radiation therapy) system from Varian medicalsystems (Varian Medical Systems, Inc., Palo Alto, Calif.). In otherembodiments, the initiation energy source 1 may be commerciallyavailable components of X-ray machines or non-medical X-ray machines.X-ray machines that produce from 10 to 150 keV X-rays are readilyavailable in the marketplace. For instance, the General ElectricDefinium series or the Siemens MULTIX series are but two examples oftypical X-ray machines designed for the medical industry, while theEagle Pack series from Smith Detection is an example of a non-medicalX-ray machine. As such, the invention is capable of performing itsdesired function when used in conjunction with commercial X-rayequipment.

In other embodiments, the initiation energy source 1 can be a radiofrequency or microwave source emitting radio waves at a frequency whichpermeates the medium and which triggers or produces secondary radiantenergy emission within the medium by interaction with the energymodulation elements 6 therein. In other embodiments, the initiationenergy source 1 can be an ultraviolet, visible, near infrared (NIR) orinfrared (IR) emitter emitting at a frequency which permeates the medium4 and which triggers or produces secondary radiant energy emissionwithin medium 4 by interaction with the energy modulation elements 6therein.

FIG. 3B is a schematic depicting another system according to anotherembodiment of the invention in which the initiation energy source 1 ofFIG. 3A is directed to energy modulation elements 6 placed in thevicinity of a fluid medium 4 (e.g., a liquid or other fluid-like medium)and held inside a container 9. The container 9 is made of a materialthat is “transparent” to the radiation 7. For example, plastic, quartz,glass, or aluminum containers would be sufficiently transparent toX-rays, while plastic or quartz or glass containers would be transparentto microwave or radio frequency light. The energy modulation elements 6can be dispersed uniformly throughout the medium or may be segregated indistinct parts of the medium or further separated physically from themedium by encapsulation structures 10. A supply 11 provides the medium 4to the container 9.

Alternatively, as shown in FIG. 3C, the luminescing particles could bepresent in the medium in encapsulated structures 10. In one embodiment,the encapsulated structures 10 are aligned with an orientation in linewith the external initiation energy source 1. In this configuration,each of the encapsulated structures 10 has itself a “line-of-sight” tothe external initiation energy source 1 shown in FIG. 3C without beingoccluded by other of the encapsulated structures 10. In otherembodiments, the encapsulated structures 10 are not so aligned in thatdirection, but could aligned perpendicular to the direction shown inFIG. 3C, or could be randomly placed. Indeed, supply of fluid medium 4could itself be used to agitate the encapsulated structures 10 and mixthe fluid medium 4 inside container 9.

The system of FIG. 3C may also be used without energy modulation agents.In this embodiment, the initiation energy source 1 can be for example atan energy suitable for driving physical, chemical, and/or biologicalprocesses in the fluid medium 4. In one aspect of the invention, theinitiation energy source 1 can a UV light source as in many conventionalUV sterilization systems and the encapsulated structures 10 of FIG. 3Care light rods conducting UV light from an exterior source to a regioninside the medium 4. In one aspect of the invention, the initiationenergy source 1 can be even disposed inside the medium and can be a UVlight source as in many conventional UV sterilization systems.

FIG. 3D is a schematic depicting a system according to anotherembodiment of the invention in which the initiation energy source isdirected a container enclosing a medium having energy modulation agentssegregated within the medium in a fluidized bed 20 configuration. Thefluidized bed 20 includes the encapsulated structures 10 in aconfiguration where a fluid to be treated is passed between theencapsulated structures 10.

In further embodiments of the invention, robotic manipulation devicesmay also be included in the systems of FIGS. 3A, 3B, 3C, and 3D for thepurpose of delivering and dispersing the energy modulation elements 6 inmedium 4 or for the purpose of removing old product and introducing newproduct for treatment into the system.

Commercial Applications

In the following commercial applications of the invention describedhere, the energy modulation agents 3 (e.g., luminescing particles orphoton emitters) are provided and distributed into a medium 4 fordeactivation or activation of agents in the medium to produce aphysical, chemical, or biological change in the medium.

Examples of luminescing particles can include gold particles (such asfor example the nanoparticles of gold described above), BaFBr:Euparticles, CdSe particles, Y₂O₃:Eu³⁺ particles, and/or other knownstimulated luminescent materials such as for example ZnS: Mn²⁺; ZnS:Mn²⁺,Yb³⁺, Y₂O₃: Eu³⁺; BaFBr:Tb³⁺; and YF₃:Tb³+.

In one embodiment of the invention described here, other potentiallyuseful luminescing particles (or energy modulation agents) includecarbon nanotubes as described for example by Wang et al. in“Electromagnetic excitation of nano-carbon in vacuum,” in OPTICSEXPRESS, Vol. 13, No. 10, May 10, 2005, the entire contents of which areincorporated herein by reference. Such carbon nanotubes show both blackbody emission and discrete line-type emissions in the visible whenexposed to microwave irradiation.

Other potentially useful luminescing particles for the inventiondescribed here include the chemiluminescent reactions/species describedby Aslan et al. in “Multicolor Microwave-Triggered Metal-EnhancedChemiluminescence,” in J. AM. CHEM. SOC. published on Web Sep. 23, 2006,the entire contents of which are incorporated herein by reference. Thesechemiluminescent reactions/species are formed with silver nanoparticleswhich enhance the chemiluminescent reactions when exposed to microwaveradiation. Aslan et al. utilized chemiluminescent species fromcommercial glow sticks where for example hydrogen peroxide oxidizesphenyl oxalate ester to a peroxyacid ester and phenol. The unstableperoxyacid ester decomposes to a peroxy compound and phenol, the processchemically inducing an electronic excited state responsible for thelight emission. While these chemiluminescent species will have a limitedlifetime, there use in curing applications for the invention describedhere is still viable where the cure process is a one-time occurrence,and the external microwave source accelerates the cure by acceleratedvisible light production.

The luminescent wavelength and/or efficiency of the luminescentparticles often depend on the size of the particle. Particle sizes inthe nanometer size range for the invention described here exhibitstronger luminescence in many cases, as described in U.S. Pat. Appl.Publ. No. 2007/0063154, whose entire contents are incorporated herein byreference. Further, in one embodiment of the invention described here,the luminescing particles can be combined with molecular complexes suchas poly(ethylene glycol), vitamin B12, or DNA, which serves to mitigateagainst coagulation of the luminescing particles (especially thenanoparticles) and serves to make the luminescing particlesbiocompatible. More specifically, one recipe for the synthesis of CdSenanocrystals is given here from U.S. Pat. Appl. Publ. No. 2007/0063154.Accordingly, citrate-stabilized CdSe nanocrystals suitable for theinvention described here can be prepared according to the followingprocedure:

-   -   To 45 ml of water are added 0.05 g sodium citrate (Fluka) and 2        ml of 4×10⁻²M cadmium perchlorate (Aldrich). The pH is adjusted        to 9.0 by 0.1 M NaOH (Alfa). The solution is bubbled with        nitrogen for 10 minutes, and then 2 ml of 1×10⁻²M        N,N-dimethylselenourea (Alfa) is added. The mixture is heated in        a conventional 900-watt microwave oven for 50 seconds. In this        recipe, the Cd:Se molar ratio is 4:1, which leads to CdSe        nanoparticles with {tilde over ( )}4.0 nm diameter; by        increasing the Cd concentration it is possible to synthesize        smaller CdSe nanoparticles.

Further, the luminescing particles for the invention described here canbe coated with insulator materials such as for example silica which willreduce the likelihood of any chemical interaction between theluminescing particles and the medium. For biological applications ofinorganic nanoparticles, one of the major limiting factors is theirtoxicity. Generally speaking, all semiconductor nanoparticles are moreor less toxic. For biomedical applications, nanoparticles with toxicityas low as possible are desirable or else the nanoparticles have toremain separated from the medium. Pure TiO₂, ZnO, and Fe₂O₃ arebiocompatible. CdTe and CdSe are toxic, while ZnS, CaS, BaS, SrS andY₂O₃ are less toxic. In addition, the toxicity of nanoparticles canresult from their inorganic stabilizers, such as TGA, or from dopantssuch as Eu²⁺, Cr³⁺ or Nd³⁺.

To reduce the toxicity or to make these nanoparticles bio-inert orbiocompatible, one embodiment of the invention described here coatsthese nanoparticles with silica. Silica is used as a coating material ina wide range of industrial colloid products from paints and magneticfluids to high-quality paper coatings. Further, silica is bothchemically and biologically inert and also is optically transparent. Inthe following recipe (from M A. Correa-Duarte, M Giesig, and L. MLiz-Marzan, Stabilization of CdS semiconductor nanoparticles againstphotodegradation by a silica coating procedure, Chem. Phys. Lett., 1998,286: 497, the entire contents of which is explicitly incorporated hereinby reference in its entirety), citrate-stabilized CdTe:Mn 2+/SiO₂nanocrystals suitable for the invention described here can be preparedwith a silica coating:

-   -   (1) To a CdTe:Mn 2+ nanoparticle solution (50 ml), a freshly        prepared aqueous solution of 3-(mercaptopropyl)trimethoxysilane        (MPS) (0.5 ml, 1 mM) (Sigma) is added under vigorous stirring.        The function of MPS is that its mercapto group can directly bond        to the surface Cd sites of CdTe, while leaving the silane groups        pointing toward solution from where silicate ions approach the        particle surface; (2) Addition of 2 ml of sodium silicate (Alfa)        solution at pH of 10.5 under vigorous stirring; (3) The        resulting dispersion (pH {tilde over ( )}8.5) is allowed to        stand for 5 days, so that silica slowly polymerizes onto the        particle surface; and (4) Transfer of the dispersion to ethanol        so that the excess dissolved silicate can precipitate out,        increasing the silica shell thickness.

Alternatively, as shown in FIG. 3C and FIG. 3D, luminescing particles inencapsulated structures 10 could be placed in the vicinity of themedium. In one embodiment for the invention described here, luminescingparticles are coated on the interior of quartz or glass tubes 9 andsealed. In another embodiment, luminescing particles could be coated onthe surface of spheres or tubes, and afterwards encapsulated with silica(or other suitable passivation layer) using a vapor deposition orsputtering process or spin-on glass process of the solution processdescribed above to make the encapsulation structures 10 which may bepart of re-entrant structures extending from walls of a container (as inFIG. 3C) or which may be part of a fluidized bed structure (as in FIG.3D).

In the either configuration, the medium to be treated would flow by theencapsulated structures 10, or flow along with encapsulated structures6, and the separation distance between the encapsulated structures 6, 10would be set a distance smaller than the UV penetration depth in themedium.

A suitable light source (such as one of the x-ray sources discussedabove) can be used to stimulate the luminescing particles in theencapsulated structures 10. In one embodiment of the invention describedhere, the concentration of luminescing particles in the medium or thespacing between the encapsulated structures 10 is set such thatluminescing particles are separated from each other in the medium byless than a UV depth of penetration into the medium. Higherconcentrations are certainly usable and will generate higher UV fluxesshould the energy source have enough intensity to “light” all theluminescing particles.

For a relatively unclouded aqueous medium, solar UV-B irradiancedecreases to 1% after penetration into the water samples between 0.2 mand 1 m, whereas UV-A penetrates on the order of several meters. Forsuch mediums, the concentration of luminescing particles is moredetermined by the time needed for the intended UV flux to producedeactivation or activation of an agent in the medium, rather than havingto be set based on a concentration of luminescent particles where themedium itself does not occlude the UV stimulated emission frompenetrating throughout the medium. The placement of the luminescentparticles in the medium and in the vicinity of the medium is notrestricted by the optical density of the medium.

Based on published data of an average of 5.2 spontaneous photons emittedfrom BaFBr:Eu²⁺ for every keV of X-ray absorbed (M. Thorns, H vonSeggern, Method for the determination of photostimulable defect centerconcentrations, production rates, and effective formation energies, J.Appl. Phys. 1994, 75: 4658-4661, the entire contents of which is hereinexplicitly incorporated by reference in its entirety.), one expects thatabout 50 photons are emitted from a CdTe nanoparticle for each 50 keVX-ray absorbed.

Based on the results in U.S. Pat. Appl. Publ. No. 2007/0063154 for X-rayspectra of CdTe/BaFBr:Eu²⁺ nanocomposites prepared using a concentrationof 0.8 ml L-cysteine stabilized CdTe particle solution in 0.2 gBaFBr:Eu²⁺ phosphor. As the X-ray irradiation time increases, the X-rayluminescence intensity of Eu²⁺ at 390 nm increases in intensity. Thisphenomenon has been discussed in W. Chen, S. P. Wang, S. Westcott, J.Zhang, A. G. Joly, and D. E. McCready, Structure and luminescence ofBaFBr. Eu ²⁺ and BaFBr: Eu ²⁺ , Tb ³⁺ phosphors and thin films, J. Appl.Phys. 2005, 97: 083506, the entire contents of these references areherein incorporated by reference in their entirety.

Hence, in one embodiment of the invention, a minimum baselineconcentration of about 10⁹ nanoparticles per cm³ for 200 nm diameterparticles is expected to be sufficient for UV emission to produce achange in the medium. The invention is not limited to this concentrationrange, but rather this range is given as an illustrative example.Indeed, higher concentrations will increase the UV emission per unittime and provide faster reactions, which in general would be consideredmore useful in industrial applications where product throughput is aconcern.

Sterilization and Cold Pasteurization of Fluids

Table 1 included below shows appropriate intensities for germicidaldestruction.

TABLE 1 Germicidal energies needed to destroy Approximate intensity(μW/cm²) required for 99% destruction of microorganisms: Bacteria 10 400Protozoa (single celled organism) 105 000  Paramecium (slipper shapedprotozoa) 200 000  Chlorella (unicellular fresh-water alga) 13 000Flagellate(protozoan or alga with flagella) 22 000 Sporozoan (parasiticprotozoans) 100 000  Virus  8 000

Accordingly, the energy modulation agents (or luminescing particles) ofthe invention (as discussed above with regard to FIGS. 3B and 3C) can beprovided on the interior of sealed quartz or glass tubes or can beprovided coated on the surface of spheres or tubes, and furtherencapsulated with a silica or passivation layer. In either configurationfor the invention described here, a medium could flow by theencapsulated structures 6, 10 with a separation distance between theencapsulated structures or the quartz or glass tubes being made smallerthan the UV penetration depth.

For example, it is known that ultraviolet (UV) with a wavelength of 254nm tends to inactivate most types of microorganisms. Most juices areopaque to UV due to the high-suspended solids in them and hence theconventional UV treatment, usually used for water treatment, cannot beused for treating juices. In order to make the process efficient, a thinfilm reactor constructed from glass has been used with the juice flowingalong the inner surface of a vertical glass tube as a thin film. See“Ultraviolet Treatment of Orange Juice” by Tran et al. published inInnovative Food Science & Emerging Technologies (Volume 5, Issue 4,December 2004, Pages 495-502), the entire contents of which areincorporated herein by reference. Tran et al. reported therein decimalreduction doses required for the reconstitute orange juices (OJ; 10.5°Brix) were 87±7 and 119±17 mJ/cm² for the standard aerobic plate count(APC) and yeast and moulds, respectively. In that article, the shelflife of fresh squeezed orange juice was extended to 5 days with alimited exposure of UV (73.8 mJ/cm²). The effect of UV on theconcentration of Vitamin C was investigated using both HPLC andtitration methods of measurements. The degradation of Vitamin C was 17%under high UV exposure of 100 mJ/cm², which was similar to that usuallyfound in thermal sterilization. Enzyme pectin methylesterase (PME)activity, which is the major cause of cloud loss of juices, was alsomeasured. The energy required for UV treatment of orange juice (2.0 kWh/m³) was much smaller than that required in thermal treatment (82 kWh/m³). The color and pH of the juice were not significantly influencedby the treatment.

The invention described herein offers advantages over this approach inthat the energy modulation agents can be placed inside fixtures such asquartz or glass (encapsulation structures 8) within the orange juice (orother fluid medium) and irradiated with x-rays (or other penetratingradiation) through for example a plastic or aluminum container 9 toactivate the energy modulation agents 3 and 6 in the orange juice. Assuch, the expense and fragility of a thin film reactor constructed fromglass of other similar structure is avoided.

While discussed with regard to orange juice, any other medium to besterilized including food products, medical products and cosmeticproducts could be treated using the technique of the invention describedherein.

Sterilization of Medical and Pharmaceutical Articles

As noted above, medical bottle caps need to be sterilized between thebase cap material and the seal material which contacts to the base ofthe medical bottle. Steam autoclaves are insufficient for this purposeas once glued, the steam is unable to penetrate into the glue seam.

Gamma irradiation has been used conventionally to sterilize medicalbottle caps and other medical, pharmaceutical, and cosmetic articlessuch as surgical disposables (e.g., surgical bandages, dressings, gaugepads, nappies, delivery kits, and etc.), metallic products (e.g.,surgical blades, implants, aluminum caps, containers, etc.), and plasticand rubber Items (e.g., petri-dish, centrifuge tube, blood collectionsets, scalp vein sets, shunt valves, rubber gloves, contraceptivedevices, gowns, wraps covers, sheets, etc.). The invention would beapplicable for the sterilization of any “interior” surfaces of these andother products.

In one embodiment of the invention described herein, UV luminescentparticles would be included in an adhesive layer when the seal materialis applied to the bottle cap. X-ray irradiation would then be capable ofcuring the adhesive (if for example the adhesive were a photosensitiveadhesive as discussed below in greater detail) and would produce withinthe adhesive medium UV radiation for direct sterilization or for theproduction of singlet oxygen or ozone for biological germicide.

While illustrated here with regard to medical bottle caps, otheradhesively constructed devices could benefit from these procedures inwhich the adhesive medium is cured and/or sterilized during activationof energy modulation agents 3 and 6.

Sterilization of Blood Products

U.S. Pat. No. 6,087,141 (the entire contents of which are incorporatedherein by reference) describes an ultraviolet light actived psoralenprocess for sterilization of blood transfusion products. Here, theinvention can be applied for example in the equipment shown in FIGS. 3Cand 3D for the treatment of or the neutralization of AIDS and HIV orother viral or pathogenic agents in blood transfusion products. In thisembodiment, at least one photoactivatable agent is selected frompsoralens, pyrene cholesteryloleate, acridine, porphyrin, fluorescein,rhodamine, 16-diazorcortisone, ethidium, transition metal complexes ofbleomycin, transition metal complexes of deglycobleomycin organoplatinumcomplexes, alloxazines, vitamin Ks, vitamin L, vitamin metabolites,vitamin precursors, naphthoquinones, naphthalenes, naphthols andderivatives thereof having planar molecular conformations,porphorinporphyrins, dyes and phenothiazine derivatives, coumarins,quinolones, quinones, and anthroquinones. These photoactivatable agentsare introduced into the blood product (or a patient's blood stream). Apenetrating energy is applied to the blood product (or to the patient).The energy modulation agents (either included in the blood product) orin encapsulated structures 10 generate secondary light such as UV lightwhich activates the photoactivatable agents in the blood products.

In a specific example, the photoactivatable agent is a psoralen, acoumarin, or a derivative thereof, and as discussed above, one cansterilize blood products in vivo (i.e., in a patient) or in a containerof the blood product (such as for example donated blood). The treatmentcan be applied to treat disorders such as for example a cancer cell, atumor cell, an autoimmune deficiency symptom virus, or a blood-bornegermicide is treated by the psoralen, the coumarin, or the derivativethereof.

As described in incorporated by reference Ser. No. 11/935,655, U.S. Pat.No. 6,235,508 further teaches that psoralens are naturally occurringcompounds which have been used therapeutically for millennia in Asia andAfrica. The action of psoralens and light has been used to treatvitiligo and psoriasis (PUVA therapy; Psoralen Ultra Violet A). Psoralenis capable of binding to nucleic acid double helices by intercalationbetween base pairs; adenine, guanine, cytosine and thymine (DNA) oruracil (RNA). Upon sequential absorption of two UV-A photons, psoralenin its excited state reacts with a thymine or uracil double bond andcovalently attaches to both strands of a nucleic acid helix. Thecrosslinking reaction appears to be specific for a thymine (DNA) or auracil (RNA) base. Binding proceeds only if psoralen is intercalated ina site containing thymine or uracil, but an initial photoadduct mustabsorb a second UVA photon to react with a second thymine or uracil onthe opposing strand of the double helix in order to crosslink each ofthe two strands of the double helix, as shown below. This is asequential absorption of two single photons as shown, as opposed tosimultaneous absorption of two or more photons.

In addition, the reference teaches that 8-MOP is unsuitable for use asan antiviral, because it damages both cells and viruses. Lethal damageto a cell or virus occurs when the psoralen is intercalated into anucleic acid duplex in sites containing two thymines (or uracils) onopposing strands but only when it sequentially absorbs 2 UVA photons andthymines (or uracils) are present. U.S. Pat. No. 4,748,120 of Wiesehanis an example of the use of certain substituted psoralens by aphotochemical decontamination process for the treatment of blood orblood products.

As described in incorporated by reference Ser. No. 11/935,655, there isprovided a novel method of treating cell proliferation disorders that iseffective, specific, and has few side-effects. Those cells sufferingfrom a cell proliferation disorder are referred to herein as the targetcells. A treatment for cell proliferation disorders, including solidtumors, is capable of chemically binding cellular nucleic acids,including but not limited to, the DNA or mitochondrial DNA or RNA of thetarget cells. For example, a photoactivatable agent, such as a psoralenor a psoralen derivative, is exposed in situ to an energy source capableof activating the photoactivatable agent or agents selected. In anotherexample, the photoactivatable agent is a photosensitizer. Thephotoactivatable agent may be a metal nanocluster or a molecule.

As noted above, an object of the present invention is to treat cellproliferation disorders. Exemplary cell proliferation disorders mayinclude, but are not limited to, cancer, as well as bacterial and viralinfections where the invading bacteria grows at a much more rapid ratethan cells of the infected host. In addition, treatment for certaindevelopmental stage diseases related to cell proliferation, such assyndactyly, are also contemplated.

Accordingly, in one embodiment, the present invention provides methodsthat are capable of overcoming the shortcomings of the existing methods.In general, a method in accordance with the present invention utilizesthe principle of energy transfer to and among molecular agents tocontrol delivery and activation of pharmaceutically active agents suchthat delivery of the desired pharmacological effect is more focused,precise, and effective than the conventional techniques.

Generally, the present invention provides methods for the treatment ofcell proliferation disorders, in which an initiation energy sourceprovides an initiation energy that activates an activatablepharmaceutical agent to treat target cells within the subject. In onepreferred embodiment, the initiation energy source is applied indirectlyto the activatable pharmaceutical agent, preferably in proximity to thetarget cells. Within the context of the present invention, the phrase“applied indirectly” (or variants of this phrase, such as “applyingindirectly”, “indirectly applies”, “indirectly applied”, “indirectlyapplying”, etc.), when referring to the application of the initiationenergy, means the penetration by the initiation energy into the subjectbeneath the surface of the subject and to the activatable pharmaceuticalagent within a subject. In one embodiment, the initiation energyinteracts with a previously administered energy modulation agent whichthen activates the activatable pharmaceutical agent. In anotherembodiment, the initiation energy itself activates the activatablepharmaceutical agent. In either embodiment, the initiation energy sourcecannot be within line-of-sight of the activatable pharmaceutical agent.By “cannot be within line-of-sight” is meant that if a hypotheticalobserver were located at the location of the activatable pharmaceuticalagent, that observer would be unable to see the source of the initiationenergy.

In one embodiment, the activation energy is capable of penetrating humantissue up to about 4 mm.

Although not intending to be bound by any particular theory or beotherwise limited in any way, the following theoretical discussion ofscientific principles and definitions are provided to help the readergain an understanding and appreciation of the present invention.

As used herein, the term “subject” is not intended to be limited tohumans, but may also include animals, plants, or any suitable biologicalorganism.

As used herein, the phrase “cell proliferation disorder” refers to anycondition where the growth rate of a population of cells is less than orgreater than a desired rate under a given physiological state andconditions. Although, preferably, the proliferation rate that would beof interest for treatment purposes is faster than a desired rate, slowerthan desired rate conditions may also be treated by methods of thepresent invention. Exemplary cell proliferation disorders may include,but are not limited to, cancer, bacterial infection, immune rejectionresponse of organ transplant, solid tumors, viral infection, autoimmunedisorders (such as arthritis, lupus, inflammatory bowel disease,Sjogrens syndrome, multiple sclerosis) or a combination thereof, as wellas aplastic conditions wherein cell proliferation is low relative tohealthy cells, such as aplastic anemia. Particularly preferred cellproliferation disorders for treatment using the present methods arecancer, staphylococcus aureus (particularly antibiotic resistant strainssuch as methicillin resistant staphylococcus aureus or MRSA), andautoimmune disorders.

As used herein, an “activatable pharmaceutical agent” is an agent thatnormally exists in an inactive state in the absence of an activationsignal. When the agent is activated by a matching activation signalunder activating conditions, it is capable of effecting the desiredpharmacological effect on a target cell (i.e. preferably a predeterminedcellular change). Signals that may be used to activate a correspondingagent may include, but are not limited to, photons of specificwavelengths (e.g. x-rays, or visible light), electromagnetic energy(e.g. radio or microwave), thermal energy, acoustic energy, or anycombination thereof. Activation of the agent may be as simple asdelivering the signal to the agent or may further premise on a set ofactivation conditions. For example, in the former case, an activatablepharmaceutical agent, such as a photosensitizer, may be activated byUV-A radiation. Once activated, the agent in its active-state may thendirectly proceed to effect a cellular change. Where activation mayfurther premise upon other conditions, mere delivery of the activationsignal may not be sufficient to bring about the desired cellular change.For example, a photoactive compound that achieves its pharmaceuticaleffect by binding to certain cellular structure in its active state mayrequire physical proximity to the target cellular structure when theactivation signal is delivered. For such activatable agents, delivery ofthe activation signal under non-activating conditions will not result inthe desired pharmacologic effect. Some examples of activating conditionsmay include, but are not limited to, temperature, pH, location, state ofthe cell, presence or absence of co-factors.

Selection of an activatable pharmaceutical agent greatly depends on anumber of factors such as the desired cellular change, the desired formof activation, as well as the physical and biochemical constraints thatmay apply. Exemplary activatable pharmaceutical agents may include, butare not limited to, agents that may be activated by photonic energy,electromagnetic energy, acoustic energy, chemical or enzymaticreactions, thermal energy, or any other suitable activation mechanisms.

When activated, the activatable pharmaceutical agent may effect cellularchanges that include, but are not limited to, apoptosis, redirection ofmetabolic pathways, up-regulation of certain genes, down-regulation ofcertain genes, secretion of cytokines, alteration of cytokine receptorresponses, or combinations thereof.

The mechanisms by which an activatable pharmaceutical agent may achieveits desired effect are not particularly limited. Such mechanisms mayinclude direct action on a predetermined target as well as indirectactions via alterations to the biochemical pathways. A preferred directaction mechanism is by binding the agent to a critical cellularstructure such as nuclear DNA, MRNA, rRNA, ribosome, mitochondrial DNA,or any other functionally important structures. Indirect mechanisms mayinclude releasing metabolites upon activation to interfere with normalmetabolic pathways, releasing chemical signals (e.g. agonists orantagonists) upon activation to alter the targeted cellular response,and other suitable biochemical or metabolic alterations.

In one preferred embodiment, the activatable pharmaceutical agent iscapable of chemically binding to the DNA or mitochondria at atherapeutically effective amount. In this embodiment, the activatablepharmaceutical agent, preferably a photoactivatable agent, is exposed insitu to an activating energy emitted from an energy modulation agent,which, in turn receives energy from an initiation energy source.

Suitable activatable agents include, but are not limited to, photoactiveagents, sono-active agents, thermo-active agents, andradio/microwave-active agents. An activatable agent may be a smallmolecule; a biological molecule such as a protein, a nucleic acid orlipid; a supramolecular assembly; a nanoparticle; or any other molecularentity having a pharmaceutical activity once activated.

The activatable agent may be derived from a natural or synthetic origin.Any such molecular entity that may be activated by a suitable activationsignal source to effect a predetermined cellular change may beadvantageously employed in the present invention.

As discussed above, suitable photoactive agents include, but are notlimited to: psoralens and psoralen derivatives, pyrenecholesteryloleate, acridine, porphyrin, fluorescein, rhodamine,16-diazorcortisone, ethidium, transition metal complexes of bleomycin,transition metal complexes of deglycobleomycin, organoplatinumcomplexes, alloxazines such as 7,8-dimethyl-10-ribityl isoalloxazine(riboflavin), 7,8,10-trimethylisoalloxazine (lumiflavin),7,8-dimethylalloxazine (lumichrome), isoalloxazine-adenine dinucleotide(flavine adenine dinucleotide [FAD]), alloxazine mononucleotide (alsoknown as flavine mononucleotide [FMN] and riboflavine-5-phosphate),vitamin Ks, vitamin L, their metabolites and precursors, andnapththoquinones, naphthalenes, naphthols and their derivatives havingplanar molecular conformations, porphyrins, dyes such as neutral red,methylene blue, acridine, toluidines, flavine (acriflavinehydrochloride) and phenothiazine derivatives, coumarins, quinolones,quinones, and anthroquinones, aluminum (111) phthalocyaninetetrasulfonate, hematoporphyrin, and phthalocyanine, and compounds whichpreferentially adsorb to nucleic acids with little or no effect onproteins. The term “alloxazine” includes isoalloxazines.

Endogenously-based derivatives include synthetically derived analogs andhomologs of endogenous photoactivated molecules, which may have or lacklower (1 to 5 carbons) alkyl or halogen substituents of thephotosensitizers from which they are derived, and which preserve thefunction and substantial non-toxicity. Endogenous molecules areinherently non-toxic and may not yield toxic photoproducts afterphotoradiation.

Table 2 lists some photoactivatable molecules capable of beingphotoactivated to induce an auto vaccine effect.

TABLE 2 SSET and TTET rate constants for bichromatic peptides η_(ex)K_(s) of K_(SSET) (S⁻¹ ) R_(model)(Å) Compound (nm) E_(SSET) donor (S⁻¹) K_(SSET) (S⁻¹ ) (Average) R₀ (Å) R (Å) (Average) E_(TTET) k_(TTET)(S⁻¹ ) 1B 224 96.3 9.5 × 10° 2.44 × 10⁸  1.87 × 10⁸ 14.7 9 9.5 266 95 1.8 × 10⁸  2.5   5 × 10² 280 94 1.36 × 10⁸  1A 224 80 9.5 × 10° 3.8 ×10⁷ 3.67 × 10⁷ 14.7 11.8 14.1 266 79 3.6 × 10⁷ 2 3.6 × 10² 280 79 3.6 ×10⁷ 2B 224 77 9.5 × 10° 3.1 × 10⁷  3.9 × 10⁷ 14.7 11.9 6.5 266 81 3.9 ×10⁷ 32 9.4 × 10³ 280 83 4.7 × 10⁷ 2A 224 69 9.5 × 10° 2.1 × 10⁷   3 ×10⁷ 14.7 12.2 8.1 74.3 5.7 × 10⁴ 266 80 3.7 × 10⁷ 280 77 3.2 × 10⁷

Table 3 lists some additional endogenous photoactivatable molecules.

TABLE 3 Bicompatible endogenous fluorophore emitters Excitation Max.Emission Max. Endogenous Fluorophores (nm) (nm) Amino acids: Tryptophan280 350 Tyrosine 275 300 Phenylalanine 260 280 Structural Proteins:Collagen 325, 360 400, 405 Elastin 290, 325 340, 400 Enzymes andCoenzymes: flavin adenine dinucleotide 450 535 reduced nicotinamidedinucelotide 290, 351 440, 400 reduced nicotinamide dinucelotide 336 464phospate Vitamins: Vitamins A 327 510 Vitamins K 335 480 Vitamins D 390480 Vitamins B₆ compounds: Pyridoxine 332, 340 460 Pyridoxamine 335 400Pyridoxal 330 385 Pyridoxic acid 315 425 Pyridoxal phosphate   5′-330400 Vitamine B₁₂ 275 305 Lipids: Phospholipids 436 540, 560 Lipofuscin340-395 540, 430-460 Ceroid 340-395 430-460, 540 Porphyrins 400-450 630,690

The nature of the predetermined cellular change will depend on thedesired pharmaceutical outcome. Exemplary cellular changes may include,but are not limited to, apoptosis, necrosis, up-regulation of certaingenes, down-regulation of certain genes, secretion of cytokines,alteration of cytokine receptor responses, or a combination thereof.

The energy modulation agent may be preferably directed to the desiredsite (e.g. a tumor) by systemic administration to a subject. Forexample, a UV-A emitting energy modulation agent may be concentrated inthe tumor site by physical insertion or by conjugating the UV-A emittingenergy modulation agent with a tumor specific carrier, such as a lipid,chitin or chitin-derivative, a chelate or other functionalized carrierthat is capable of concentrating the UV-A emitting source in a specifictarget tumor. In one embodiment, the energy modulation agent is a singleenergy modulation agent, and is coupled to at least one activatablepharmaceutical agent.

Additionally, as above, the energy modulation agent can be used alone oras a series of two or more energy modulation agents wherein the energymodulation agents provide an energy cascade. Thus, the first energymodulation agent in the cascade will absorb the activation energy,convert it to a different energy which is then absorbed by the secondenergy modulation in the cascade, and so forth until the end of thecascade is reached with the final energy modulation agent in the cascadeemitting the energy necessary to activate the activatable pharmaceuticalagent.

As above, although the activatable pharmaceutical agent and the energymodulation agent can be distinct and separate, it will be understoodthat the two agents need not be independent and separate entities. Infact, the two agents may be associated with each other via a number ofdifferent configurations. Where the two agents are independent andseparately movable from each other, they generally interact with eachother via diffusion and chance encounters within a common surroundingmedium. Where the activatable pharmaceutical agent and the energymodulation agent are not separate, they may be combined into one singleentity.

The initiation energy source can be any energy source capable ofproviding energy at a level sufficient to activate the activatable agentdirectly, or to provide the energy modulation agent with the inputneeded to emit the activation energy for the activatable agent (indirectactivation). Preferable initiation energy sources include, but are notlimited to, UV-A lamps or fiber optic lines, a light needle, anendoscope, and a linear accelerator that generates x-ray, gamma-ray, orelectron beams. In a preferred embodiment the initiation energy capableof penetrating completely through the subject. Within the context of thepresent invention, the phrase “capable of penetrating completely throughthe subject” is used to refer to energy that can penetrate to any depthwithin the subject to activate the activatable pharmaceutical agent. Itis not required that the any of the energy applied actually passcompletely through the subject, merely that it be capable of doing so inorder to permit penetration to any desired depth to activate theactivatable pharmaceutical agent. Exemplary initiation energy sourcesthat are capable of penetrating completely through the subject include,but are not limited to, x-rays, gamma rays, electron beams, microwavesand radio waves.

In one embodiment, the source of the initiation energy can be aradiowave emitting nanotube, such as those described by K. Jensen, J.Weldon, H. Garcia, and A. Zettl in the Department of Physics at theUniversity of California at Berkeley (seehttp://socrates.berkeley.edu/{tilde over ( )}argon/nanoradio/radio.html,the entire contents of which are hereby incorporated by reference).These nanotubes can be administered to the subject, and preferably wouldbe coupled to the activatable pharmaceutical agent or the energymodulation agent, or both, such that upon application of the initiationenergy, the nanotubes would accept the initiation energy (preferablyradiowaves), then emit radiowaves in close proximity to the activatablepharmaceutical agent, or in close proximity to the energy modulationagent, to then cause activation of the activatable pharmaceutical agent.In such an embodiment, the nanotubes would act essentially as aradiowave focusing or amplification device in close proximity to theactivatable pharmaceutical agent or energy modulation agent.

Signal transduction schemes as a drug delivery vehicle may beadvantageously developed by careful modeling of the cascade eventscoupled with metabolic pathway knowledge to sequentially orsimultaneously activate multiple activatable pharmaceutical agents toachieve multiple-point alterations in cellular function.

Photoactivatable agents may be stimulated by an energy source, such asirradiation, resonance energy transfer, exciton migration, electroninjection, or chemical reaction, to an activated energy state that iscapable of effecting the predetermined cellular change desired. In apreferred embodiment, the photoactivatable agent, upon activation, bindsto DNA or RNA or other structures in a cell. The activated energy stateof the agent is capable of causing damage to cells, inducing apoptosis.The mechanism of apoptosis is associated with an enhanced immuneresponse that reduces the growth rate of cell proliferation disordersand may shrink solid tumors, depending on the state of the patient'simmune system, concentration of the agent in the tumor, sensitivity ofthe agent to stimulation, and length of stimulation.

A preferred method of treating a cell proliferation disorder of thepresent invention administers a photoactivatable agent to a patient,stimulates the photoactivatable agent to induce cell damage, andgenerates an auto vaccine effect. In one further preferred embodiment,the photoactivatable agent is stimulated via a resonance energytransfer.

One advantage is that multiple wavelengths of emitted radiation may beused to selectively stimulate one or more photoactivatable agents orenergy modulation agents capable of stimulating the one or morephotoactivatable agents. The energy modulation agent is preferablystimulated at a wavelength and energy that causes little or no damage tohealthy cells, with the energy from one or more energy modulation agentsbeing transferred, such as by Foerster Resonance Energy Transfer, to thephotoactivatable agents that damage the cell and cause the onset of thedesired cellular change, such as apoptosis of the cells.

Another advantage is that side effects can be greatly reduced bylimiting the production of free radicals, singlet oxygen, hydroxides andother highly reactive groups that are known to damage healthy cells.Furthermore, additional additives, such as antioxidants, may be used tofurther reduce undesired effects of irradiation.

As noted above, Resonance Energy Transfer (RET) is an energy transfermechanism between two molecules having overlapping emission andabsorption bands. Electromagnetic emitters are capable of converting anarriving wavelength to a longer wavelength. For example, UV-B energyabsorbed by a first molecule may be transferred by a dipole-dipoleinteraction to a UV-A-emitting molecule in close proximity to theUV-B-absorbing molecule. Alternatively, a material absorbing a shorterwavelength may be chosen to provide RET to a non-emitting molecule thathas an overlapping absorption band with the transferring molecule'semission band. Alternatively, phosphorescence, chemiluminescence, orbioluminescence may be used to transfer energy to a photoactivatablemolecule.

Alternatively, one can administer the initiation energy source to thesubject. Within the context of the present invention, the administeringof the initiation energy source means the administration of an agent,that itself produces the initiation energy, in a manner that permits theagent to arrive at the target cell within the subject without beingsurgically inserted into the subject. The administration can take anyform, including, but not limited to, oral, intravenous, intraperitoneal,inhalation, etc. Further, the initiation energy source in thisembodiment can be in any form, including, but not limited to, tablet,powder, liquid solution, liquid suspension, liquid dispersion, gas orvapor, etc. In this embodiment, the initiation energy source includes,but is not limited to, chemical energy sources, nanoemitters, nanochips,and other nanomachines that produce and emit energy of a desiredfrequency. Recent advances in nanotechnology have provided examples ofvarious devices that are nanoscale and produce or emit energy, such asthe Molecular Switch (or Mol-Switch) work by Dr. Keith Firman of the ECResearch and Development Project, or the work of Cornell et al. (1997)who describe the construction of nanomachines based around ion-channelswitches only 1.5 nm in size, which use ion channels formed in anartificial membrane by two gramicidin molecules: one in the lower layerof the membrane attached to a gold electrode and one in the upper layertethered to biological receptors such as antibodies or nucleotides. Whenthe receptor captures a target molecule or cell, the ion channel isbroken, its conductivity drops, and the biochemical signal is convertedinto an electrical signal. These nanodevices could also be coupled withthe present invention to provide targeting of the target cell, todeliver the initiation energy source directly at the desired site. Inanother embodiment, the present invention includes the administration ofthe activatable pharmaceutical agent, along with administration of asource of chemical energy such as chemiluminescence, phosphorescence orbioluminescence. The source of chemical energy can be a chemicalreaction between two or more compounds, or can be induced by activatinga chemiluminescent, phosphorescent or bioluminescent compound with anappropriate activation energy, either outside the subject or inside thesubject, with the chemiluminescence, phosphorescence or bioluminescencebeing allowed to activate the activatable pharmaceutical agent in vivoafter administration. The administration of the activatablepharmaceutical agent and the source of chemical energy can be performedsequentially in any order or can be performed simultaneously. In thecase of certain sources of such chemical energy, the administration ofthe chemical energy source can be performed after activation outside thesubject, with the lifetime of the emission of the energy being up toseveral hours for certain types of phosphorescent materials for example.There are no known previous efforts to use resonance energy transfer ofany kind to activate an intercalator to bind DNA.

Yet another example is that nanoparticles or nanoclusters of certainatoms may be introduced such that are capable of resonance energytransfer over comparatively large distances, such as greater than onenanometer, more preferably greater than five nanometers, even morepreferably at least 10 nanometers. Functionally, resonance energytransfer may have a large enough “Foerster” distance (R₀), such thatnanoparticles in one part of a cell are capable of stimulatingactivation of photoactivatable agents disposed in a distant portion ofthe cell, so long as the distance does not greatly exceed R₀. Forexample, gold nanospheres having a size of 5 atoms of gold have beenshown to have an emission band in the ultraviolet range, recently.

The present invention treatment may also be used for inducing an autovaccine effect for malignant cells, including those in solid tumors. Tothe extent that any rapidly dividing cells or stem cells may be damagedby a systemic treatment, then it may be preferable to direct thestimulating energy directly toward the tumor, preventing damage to mostnormal, healthy cells or stem cells by avoiding photoactivation orresonant energy transfer of the photoactivatable agent.

Alternatively, a treatment may be applied that slows or pauses mitosis.Such a treatment is capable of slowing the division of rapidly dividinghealthy cells or stem cells during the treatment, without pausingmitosis of cancerous cells. Alternatively, a blocking agent isadministered preferentially to malignant cells prior to administeringthe treatment that slows mitosis.

In one embodiment, an aggressive cell proliferation disorder has a muchhigher rate of mitosis, which leads to selective destruction of adisproportionate share of the malignant cells during even a systemicallyadministered treatment. Stem cells and healthy cells may be spared fromwholesale programmed cell death, even if exposed to photoactivatedagents, provided that such photoactivated agents degenerate from theexcited state to a lower energy state prior to binding, mitosis or othermechanisms for creating damage to the cells of a substantial fraction ofthe healthy stem cells. Thus, an auto-immune response may not beinduced.

Alternatively, a blocking agent may be used that prevents or reducesdamage to stem cells or healthy cells, selectively, which wouldotherwise be impaired. The blocking agent is selected or is administeredsuch that the blocking agent does not impart a similar benefit tomalignant cells, for example.

In one embodiment, stem cells are targeted, specifically, fordestruction with the intention of replacing the stem cells with a donorcell line or previously stored, healthy cells of the patient. In thiscase, no blocking agent is used. Instead, a carrier or photosensitizeris used that specifically targets the stem cells.

Any of the photoactivatable agents may be exposed to an excitationenergy source implanted in a tumor. The photoactive agent may bedirected to a receptor site by a carrier having a strong affinity forthe receptor site. Within the context of the present invention, a“strong affinity” is preferably an affinity having an equilibriumdissociation constant, K_(i), at least in the nanomolar, nM, range orhigher. Preferably, the carrier may be a polypeptide and may form acovalent bond with a photoactive agent, for example. The polypeptide maybe an insulin, interleukin, thymopoietin or transferrin, for example.Alternatively, a photoactive agent may have a strong affinity for thetarget cell without binding to a carrier.

A receptor site may be any of the following: nucleic acids of nucleatedblood cells, molecule receptor sites of nucleated blood cells, theantigenic sites on nucleated blood cells, epitopes, or other sites wherephotoactive agents are capable of destroying a targeted cell.

In one embodiment, thin fiber optic lines are inserted in the tumor andlaser light is used to photoactivate the agents. In another embodiment,a plurality of sources for supplying electromagnetic radiation energy orenergy transfer are provided by one or more molecules administered to apatient. The molecules may emit stimulating radiation in the correctband of wavelength to stimulate the photoactivatable agents, or themolecules may transfer energy by a resonance energy transfer or othermechanism directly to the photoactivatable agent or indirectly by acascade effect via other molecular interactions.

In another embodiment, the patient's own cells are removed andgenetically modified to provide photonic emissions. For example, tumoror healthy cells may be removed, genetically modified to inducebioluminescence and may be reinserted at the site of the tumor to betreated. The modified, bioluminescent cells may be further modified toprevent further division of the cells or division of the cells only solong as a regulating agent is present. Administration of anintercalator, systemically or targeting tumor cells, that is capable ofphotoactivation by bioluminescent cells may produce conditions suitablefor creating an auto vaccine effect due to apoptosis of malignant cells.Preferably, apoptosis triggers and stimulates the body to develop animmune response targeting the malignant cells.

Similar to that noted above, in a further embodiment, a biocompatibleemitting source, such as a fluorescing metal nanoparticle or fluorescingdye molecule, is selected that emits in the UV-A band. The UV-A emittingsource is directed to the site of a tumor. The UV-A emitting source maybe directed to the site of the tumor by systemically administering theUV-A emitting source. Preferably, the UV-A emitting source isconcentrated in the tumor site, such as by physical insertion or byconjugating the UV-A emitting molecule with a tumor specific carrier,such as a lipid, chitin or chitin-derivative, a chelate or otherfunctionalized carrier that is capable of concentrating the UV-Aemitting source in a specific target tumor, as is known in the art.

Similar to that noted above, in one preferred embodiment, the UV-Aemitting source is a gold nanoparticle comprising a cluster of 5 goldatoms, such as a water soluble quantum dot encapsulated bypolyamidoamine dendrimers. The gold atom clusters may be producedthrough a slow reduction of gold salts (e.g. HAuCl₄ or AuBr₃) or otherencapsulating amines, for example. One advantage of such a goldnanoparticle is the increased Foerster distance (i.e. R₀), which may begreater than 100 angstroms. The equation for determining the Foersterdistance is substantially different from that for molecularfluorescence, which is limited to use at distances less than 100angstroms. It is believed that the gold nanoparticles are governed bynanoparticle surface to dipole equations with a 1/R⁴ distance dependencerather than a 1/R⁶ distance dependence. For example, this permitscytoplasmic to nuclear energy transfer between metal nanoparticles and aphotoactivatable molecule, such as a psoralen and more preferably an8-methoxypsoralen (8-MOP) administered orally to a patient, which isknown to be safe and effective at inducing an apoptosis of leukocytes.

Similar to that noted above, in another embodiment, a UV- orlight-emitting luciferase is selected as the emitting source forexciting a photoactivatable agent. A luciferase may be combined with ATPor another molecule, which may then be oxygenated with additionalmolecules to stimulate light emission at a desired wavelength.Alternatively, a phosphorescent emitting source may be used. Oneadvantage of a phosphorescent emitting source is that the phosphorescentemitting molecules or other source may be electroactivated orphotoactivated prior to insertion into the tumor either by systemicadministration or direct insertion into the region of the tumor.Phosphorescent materials may have longer relaxation times thanfluorescent materials, because relaxation of a triplet state is subjectto forbidden energy state transitions, storing the energy in the excitedtriplet state with only a limited number of quantum mechanical energytransfer processes available for returning to the lower energy state.Energy emission is delayed or prolonged from a fraction of a second toseveral hours. Otherwise, the energy emitted during phosphorescentrelaxation is not otherwise different than fluorescence, and the rangeof wavelengths may be selected by choosing a particular phosphor.

Similar to that noted above, in another embodiment, a combinedelectromagnetic energy harvester molecule is designed, such as thecombined light harvester disclosed in J. Am. Chem. Soc. 2005, 127,9760-9768, the entire contents of which are hereby incorporated byreference. By combining a group of fluorescent molecules in a molecularstructure, a resonance energy transfer cascade may be used to harvest awide band of electromagnetic radiation resulting in emission of a narrowband of fluorescent energy. By pairing a combined energy harvester witha photoactivatable molecule, a further energy resonance transfer excitesthe photoactivatable molecule, when the photoactivatable molecule isnearby stimulated combined energy harvester molecules. Another exampleof a harvester molecule is disclosed in FIG. 4 of “Singlet-Singlet andTriplet-Triplet Energy Transfer in Bichromophoric Cyclic Peptides,” M.S. Thesis by M. O. Guler, Worcester Polytechnic Institute, May 18, 2002,which is incorporated herein by reference.

Similar to that noted above, in another embodiment, a Stokes shift of anemitting source or a series of emitting sources arranged in a cascade isselected to convert a shorter wavelength energy, such as X-rays, to alonger wavelength fluorescence emission such a optical or UV-A, which isused to stimulate a photoactivatable molecule at the location of thetumor cells. Preferably, the photoactivatable molecule is selected tocause an apoptosis sequence in tumor cells without causing substantialharm to normal, healthy cells. More preferably, the apoptosis sequencethen leads to an auto vaccine effect that targets the malignant tumorcells throughout the patient's body.

Similar to that noted above, in an additional embodiment, thephotoactivatable agent can be a photocaged complex having an activeagent (which can be a cytotoxic agent or can be an activatablepharmaceutical agent) contained within a photocage. The active agent isbulked up with other molecules that prevent it from binding to specifictargets, thus masking its activity. When the photocage complex isphotoactivated (e.g., by UVA), the bulk falls off, exposing the activeagent. In such a photocage complex, the photocage molecules can bephotoactive (i.e. when photoactivated, they are caused to dissociatefrom the photocage complex, thus exposing the active agent within), orthe active agent can be the photoactivatable agent (which whenphotoactivated causes the photocage to fall off), or both the photocageand the active agent are photoactivated, with the same or differentwavelengths. For example, a toxic chemotherapeutic agent can bephotocaged, which will reduce the systemic toxicity when delivered. Oncethe agent is concentrated in the tumor, the agent is irradiated with anactivation energy. This causes the “cage” to fall off, leaving acytotoxic agent in the tumor cell. Suitable photocages include thosedisclosed by Young and Deiters in “Photochemical Control of BiologicalProcesses”, Org. Biomol. Chem., 5, pp. 999-1005 (2007) and“Photochemical Hammerhead Ribozyme Activation”, Bioorganic &MedicinalChemistry Letters, 16(10), pp. 2658-2661 (2006), the contents of whichare hereby incorporated by reference.

Similar to that noted above, in a further embodiment, some of the tumorcells are treated in vitro using a UV-A source to stimulate 8-MOP.Apoptosis of the tumor cells is monitored, and some or all of thefragments and remnants of the apoptosis process are reintroduced intothe site of a tumor. Preferably, the portion of fragments, cellularstructures and remnants are selected such that an auto vaccine effect isgenerated that leads to further apoptosis of tumor cells withoutsubstantially harming healthy tissues, causing solid tumors to shrink.

Similar to that noted above, in one embodiment, a lanthanide chelatecapable of intense luminescence is used. For example, a lanthanidechelator may be covalently joined to a coumarin or coumarin derivativeor a quinolone or quinolone-derivative sensitizer. Sensitizers may be a2- or 4-quinolone, a 2- or 4-coumarin, or derivatives or combinations ofthese examples. A carbostyril 124 (7-amino-4-methyl-2-quinolone), acoumarin 120 (7-amino-4-methyl-2-coumarin), a coumarin 124(7-amino-4-(trifluoromethyl)-2-coumarin), aminoinethyltrimethylpsoralenor other similar sensitizer may be used. Chelates may be selected toform high affinity complexes with lanthanides, such as terbium oreuropium, through chelator groups, such as DTPA. Such chelates may becoupled to any of a wide variety of well known probes or carriers, andmay be used for resonance energy transfer to a psoralen orpsoralen-derivative, such as 8-MOP, or other photoactive moleculescapable of binding DNA (i.e., a DNA intercalator) and causing theinitiation of an apoptosis process of rapidly dividing cancer cells. Inthis way, the treatment may be targeted to especially aggressive formsof cell proliferation disorders that are not successfully treated byconventional chemotherapy, radiation or surgical techniques. In onealternative example, the lanthanide chelate is localized at the site ofthe tumor using an appropriate carrier molecule, particle or polymer,and a source of electromagnetic energy is introduced by minimallyinvasive procedures to irradiate the tumor cells, after exposure to thelanthanide chelate and a photoactive molecule.

Similar to that noted above, in another embodiment, a biocompatible,endogenous fluorophore emitter is selected to stimulate resonance energytransfer to a photoactivatable molecule. A biocompatible emitter with anemission maxima within the absorption range of the biocompatible,endogenous fluorophore emitter may be selected to stimulate an excitedstate in fluorophore emitter. One or more halogen atoms may be added toany cyclic ring structure capable of intercalation between the stackednucleotide bases in a nucleic acid (either DNA or RNA) to confer newphotoactive properties to the intercalator. Any intercalating molecule(psoralens, coumarins, or other polycyclic ring structures) may beselectively modified by halogenation or addition of non-hydrogen bondingionic substituents to impart advantages in its reaction photochemistryand its competitive binding affinity for nucleic acids over cellmembranes or charged proteins, as is known in the art.

Recently, photosensitizers have been developed for treating cellproliferation disorders using photodynamic therapy. Table 4 provides anassortment of known photosensitizers that are useful in treating cellproliferation disorders.

TABLE 4 Photosensitizers for cell proliferation disorders Length ofPhoto- Drug-light Wavelength photo- sensitizer Dose interval ofactivation sensitization Photofrin 2 mg/kg 48 hrs 630 nm 4-6 weeks (II)Foscan 0.1 mg/kg 4-6 days 652 nm 2 weeks Lutatium 2-6 mg/kg 3 to 24 hrs732 nm 24-48 hrs texahyrin

Skin photosensitivity is a major toxicity of the photosensitizers.Severe sunburn occurs if skin is exposed to direct sunlight for even afew minutes. Early murine research hinted at a vigorous and long termstimulation of immune response; however, actual clinical testing hasfailed to achieve the early promises of photodynamic therapies. Theearly photosensitizers for photodynamic therapies targeted type IIresponses, which created singlet oxygen when photoactivated in thepresence of oxygen. The singlet oxygen caused cellular necrosis and wasassociated with inflammation and an immune response. However, tumors arenow known to down regulate the immune response over time, and it isthought that this is one of the reasons that clinical results are not asdramatic as promised by the early murine research. Some additionalphotosensitizers have been developed to induce type I responses,directly damaging cellular structures, which result in apoptosis oftumor cells.

Porfimer sodium (Photofrin; QLT Therapeutics, Vancouver, BC, Canada), isa partially purified preparation of hematoporphyrin derivative (HpD).Photofrin has been approved by the US Food and Drug Administration forthe treatment of obstructing esophageal cancer, microinvasiveendobronchial non-small cell lung cancer, and obstructing endobronchialnon-small cell lung cancer. Photofrin is activated with 630 nm, whichhas a tissue penetration of approximately 2 to 5 mm. Photofrin has arelatively long duration of skin photosensitivity (approximately 4 to 6weeks).

Tetra(m-hydroxyphenyl)chlorin (Foscan; Scotia Pharmaceuticals, Stirling,UK), is a synthetic chlorin compound that is activated by 652 nm light.Clinical studies have demonstrated a tissue effect of up to 10 mm withFoscan and 652 nm light. Foscan is more selectively a photosensitizer intumors than normal tissues, and requires a comparatively short lightactivation time. A recommended dose of 0.1 mg/kg is comparatively lowand comparatively low doses of light may be used. Nevertheless, durationof skin photosensitivity is reasonable (approximately 2 weeks). However,Foscan induces a comparatively high yield of singlet oxygen, which maybe the primary mechanism of DNA damage for this molecule.

Motexafin lutetium (Lutetium texaphyrin) is activated by light in thenear infrared region (732 nm). Absorption at this wavelength has theadvantage of potentially deeper penetration into tissues, compared withthe amount of light used to activate other photosensitizers (FIGS. 3Eand 3F). Lutetium texaphryin also has one of the greatest reportedselectivities for tumors compared to selectivities of normal tissues.Young S W, et al.: Lutetium texaphyrin (PCI-0123) a near-infrared,water-soluble photosensitizer. Photochem Photobiol 1996, 63:892-897. Inaddition, its clinical use is associated with a shorter duration of skinphotosensitivity (24 to 48 hours). Lutetium texaphryin has beenevaluated for metastatic skin cancers. It is currently underinvestigation for treatment of recurrent breast cancer and for locallyrecurrent prostate cancer. The high selectivity for tumors promisesimproved results in clinical trials.

In general, the approach may be used with any source for the excitationof higher electronic energy states, such as electrical, chemical and/orradiation, individually or combined into a system for activating anactivatable molecule. The process may be a photophoresis process or maybe similar to photophoresis. While photophoresis is generally thought tobe limited to photonic excitation, such as by UV-light, other forms ofradiation may be used as a part of a system to activate an activatablemolecule. Radiation includes ionizing radiation which is high energyradiation, such as an X-ray or a gamma ray, which interacts to produceion pairs in matter. Radiation also includes high linear energy transferirradiation, low linear energy transfer irradiation, alpha rays, betarays, neutron beams, accelerated electron beams, and ultraviolet rays.Radiation also includes proton, photon and fission-spectrum neutrons.Higher energy ionizing radiation may be combined with chemical processesto produce energy states favorable for resonance energy transfer, forexample. Other combinations and variations of these sources ofexcitation energy may be combined as is known in the art, in order tostimulate the activation of an activatable molecule, such as 8-MOP. Inone example, ionizing radiation is directed at a solid tumor andstimulates, directly or indirectly, activation of 8-MOP, as well asdirectly damaging the DNA of malignant tumor cells. In this example,either the effect of ionizing radiation or the photophoresis-likeactivation of 8-MOP may be thought of as an adjuvant therapy to theother.

In one embodiment, the present invention provides a method for treatinga cell proliferation disorder in a subject, comprising:

-   -   (1) administering to the subject an activatable pharmaceutical        agent that is capable of effecting a predetermined cellular        change when activated; and    -   (2) applying an initiation energy from an initiation energy        source to the subject,        wherein the initiation energy source is a source of energy        capable of penetrating completely through the subject, and        wherein the applying activates the activatable agent in situ,

thus causing the predetermined cellular change to occur, whereinoccurrence of the predetermined cellular change causes an increase inrate or decrease in rate of cell proliferation to treat the cellproliferation disorder.

In a further embodiment, the present invention provides a method fortreating a cell proliferation disorder in a subject, comprising:

(1) administering to the subject one or more energy modulation agentsand an activatable pharmaceutical agent that is capable of effecting apredetermined cellular change when activated; and

(2) applying an initiation energy from an initiation energy source tothe subject, wherein the one or more energy modulation agents convertthe initiation energy applied to UV-A or visible energy, which thenactivates the activatable agent in situ,

thus causing the predetermined cellular change to occur, whereinoccurrence of the predetermined cellular change causes an increase inrate or decrease in rate of cell proliferation to treat the cellproliferation disorder.

In a further embodiment, the present invention provides a method fortreating a cell proliferation disorder in a subject, comprising:

(1) administering to the subject an activatable pharmaceutical agentthat is capable of effecting a predetermined cellular change whenactivated; and

(2) applying an initiation energy from an initiation energy source tothe subject,

wherein the initiation energy applied and activatable pharmaceuticalagent upon activation produce insufficient singlet oxygen in the subjectto produce cell lysis, and wherein the initiation energy activates theactivatable pharmaceutical agent in situ,

thus causing the predetermined cellular change to occur, whereinoccurrence of the predetermined cellular change causes an increase inrate or decrease in rate of cell proliferation to treat the cellproliferation disorder.

Similar to that noted above, work in the area of photodynamic therapyhas shown that the amount of singlet oxygen required to cause celllysis, and thus cell death, is 0.32×10⁻³ mol/liter or more, or 10⁹singlet oxygen molecules/cell or more. However, in the presentinvention, it is most preferable to avoid production of an amount ofsinglet oxygen that would cause cell lysis, due to its indiscriminatenature of attack, lysing both target cells and healthy cells.Accordingly, it is most preferred in the present invention that thelevel of singlet oxygen production caused by the initiation energy usedor activatable pharmaceutical agent upon activation be less than levelneeded to cause cell lysis.

In yet another embodiment, the activatable pharmaceutical agent,preferably a photoactive agent, is directed to a receptor site by acarrier having a strong affinity for the receptor site. The carrier maybe a polypeptide and may form a covalent bond with a photo active agent,for example. The polypeptide may be an insulin, interleukin,thymopoietin or transferrin, for example. Alternatively, a photoactivepharmaceutical agent may have a strong affinity for the target cellwithout a binding to a carrier (i.e., a non-covalent bond).

For example, a treatment may be applied that acts to slow or pausemitosis. Such a treatment is capable of slowing the division of rapidlydividing healthy cells or stem cells without pausing mitosis ofcancerous cells. Thus, the difference in growth rate between thenon-target cells and target cells are further differentiated to enhancethe effectiveness of the methods of the present invention.

In another example, an aggressive cell proliferation disorder has a muchhigher rate of mitosis, which leads to selective destruction of adisproportionate share of the malignant cells during even a systemicallyadministered treatment. Stem cells and healthy cells may be spared fromwholesale programmed cell death even if exposed to photoactivated agentsthat cause apoptosis, provided that such photoactivated agentsdegenerate from the excited state to a lower energy state prior tobinding, mitosis or other mechanisms for creating damage to the cells ofa substantial fraction of the healthy stem cells. To further protecthealthy cells from the effect of photoactivatable agents, blockingagents that block uptake of the photoactivatable agents, prior to theiractivation, may be administered.

U.S. Pat. No. 6,235,508, discloses that a variety of blocking agentshave been found to be suitable for this purpose, some of which aretraditional antioxidants, and some of which are not. Suitable blockingagents include, but are not limited to, histidine, cysteine, tryrosine,tryptophan, ascorbate, N-acetyl cysteine, propyl gallate,mercaptopropionyl glycine, butylated hydroxytoluene (BHT) and butylatedhydroxyanisole (BHA).

In a further embodiment, methods in accordance with the presentinvention may further include adding an additive to alleviate treatmentside-effects. Exemplary additives may include, but are not limited to,antioxidants, adjuvant, or combinations thereof. In one exemplaryembodiment, psoralen is used as the activatable pharmaceutical agent,UV-A is used as the activating energy, and antioxidants are added toreduce the unwanted side-effects of irradiation.

The activatable pharmaceutical agent and derivatives thereof as well asthe energy modulation agent, can be incorporated into pharmaceuticalcompositions suitable for administration. Such compositions typicallycomprise the activatable pharmaceutical agent and a pharmaceuticallyacceptable carrier. The pharmaceutical composition also comprises atleast one additive having a complementary therapeutic or diagnosticeffect, wherein the additive is one selected from an antioxidant, anadjuvant, or a combination thereof.

As used herein, “pharmaceutically acceptable carrier” is intended toinclude any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents, and thelike, compatible with pharmaceutical administration. The use of suchmedia and agents for pharmaceutically active substances is well known inthe art. Except insofar as any conventional media or agent isincompatible with the active compound, use thereof in the compositionsis contemplated. Supplementary active compounds can also be incorporatedinto the compositions. Modifications can be made to the compound of thepresent invention to affect solubility or clearance of the compound.These molecules may also be synthesized with D-amino acids to increaseresistance to enzymatic degradation. If necessary, the activatablepharmaceutical agent can be co-administered with a solubilizing agent,such as cyclodextran.

A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (topical),transmucosal, rectal administration, and direct injection into theaffected area, such as direct injection into a tumor. Solutions orsuspensions used for parenteral, intradermal, or subcutaneousapplication can include the following components: a sterile diluent suchas water for injection, saline solution, fixed oils, polyethyleneglycols, glycerin, propylene glycol or other synthetic solvents;antibacterial agents such as benzyl alcohol or methyl parabens;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as ethylenediaminetetraacetic acid; buffers such as acetates,citrates or phosphates, and agents for the adjustment of tonicity suchas sodium chloride or dextrose. The pH can be adjusted with acids orbases, such as hydrochloric acid or sodium hydroxide. The parenteralpreparation can be enclosed in ampoules, disposable syringes or multipledose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, orphosphate buffered saline (PBS). In all cases, the composition must besterile and should be fluid to the extent that easy syringabilityexists. It must be stable under the conditions of manufacture andstorage and must be preserved against the contaminating action ofmicroorganisms such as bacteria and fungi. The carrier can be a solventor dispersion medium containing, for example, water, ethanol, polyol(for example, glycerol, propylene glycol, and liquid polyethyleneglycol, and the like), and suitable mixtures thereof. The properfluidity can be maintained, for example, by the use of a coating such aslecithin, by the maintenance of the required particle size in the caseof dispersion and by the use of surfactants. Prevention of the action ofmicroorganisms can be achieved by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, ascorbic acid,thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, polyalcohols such asmanitol, sorbitol, sodium chloride in the composition. Prolongedabsorption of the injectable compositions can be brought about byincluding in the composition an agent which delays absorption, forexample, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the activecompound in the required amount in an appropriate solvent with one or acombination of ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the active compound into a sterile vehicle that contains abasic dispersion medium and the required other ingredients from thoseenumerated above. In the case of sterile powders for the preparation ofsterile injectable solutions, methods of preparation are vacuum dryingand freeze-drying that yields a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

Oral compositions generally include an inert diluent or an ediblecarrier. They can be enclosed in gelatin capsules or compressed intotablets. For the purpose of oral therapeutic administration, the activecompound can be incorporated with excipients and used in the form oftablets, troches, or capsules. Oral compositions can also be preparedusing a fluid carrier for use as a mouthwash, wherein the compound inthe fluid carrier is applied orally and swished and expectorated orswallowed. Pharmaceutically compatible binding agents, and/or adjuvantmaterials can be included as part of the composition. The tablets,pills, capsules, troches and the like can contain any of the followingingredients, or compounds of a similar nature: a binder such asmicrocrystalline cellulose, gum tragacanth or gelatin; an excipient suchas starch or lactose, a disintegrating agent such as alginic acid,Primogel, or corn starch; a lubricant such as magnesium stearate orSterotes; a glidant such as colloidal silicon dioxide; a sweeteningagent such as sucrose or saccharin; or a flavoring agent such aspeppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in theform of an aerosol spray from pressured container or dispenser whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art, and include, forexample, for transmucosal administration, detergents, bile salts, andfusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the active compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

The compounds can also be prepared in the form of suppositories (e.g.,with conventional suppository bases such as cocoa butter and otherglycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially.Liposomal suspensions (including liposomes targeted to infected cellswith monoclonal antibodies to viral antigens) can also be used aspharmaceutically acceptable carriers. These can be prepared according tomethods known to those skilled in the art, for example, as described inU.S. Pat. No. 4,522,811.

It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved, and the limitationsinherent in the art of compounding such an active compound for thetreatment of individuals.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

Methods of administering agents according to the present invention arenot limited to the conventional means such as injection or oralinfusion, but include more advanced and complex forms of energytransfer. For example, genetically engineered cells that carry andexpress energy modulation agents may be used. Cells from the host may betransfected with genetically engineered vectors that expressbioluminescent agents. Transfection may be accomplished via in situ genetherapy techniques such as injection of viral vectors or gene guns, ormay be performed ex vivo by removing a sample of the host's cells andthen returning to the host upon successful transfection.

Such transfected cells may be inserted or otherwise targeted at the sitewhere diseased cells are located. In this embodiment, the initiationenergy source may be a biochemical source as such ATP, in which case theinitiation energy source is considered to be directly implanted in thetransfected cell. Alternatively, a conventional micro-emitter devicecapable of acting as an initiation energy source may be transplanted atthe site of the diseased cells.

It will also be understood that the order of administering the differentagents is not particularly limited. Thus in some embodiments theactivatable pharmaceutical agent may be administered before the energymodulation agent, while in other embodiments the energy modulation agentmay be administered prior to the activatable pharmaceutical agent. Itwill be appreciated that different combinations of ordering may beadvantageously employed depending on factors such as the absorption rateof the agents, the localization and molecular trafficking properties ofthe agents, and other pharmacokinetics or pharmacodynamicsconsiderations.

An advantage of the methods of the present invention is that byspecifically targeting cells affected by a cell proliferation disorder,such as rapidly dividing cells, and triggering a cellular change, suchas apoptosis, in these cells in situ, the immune system of the host maybe stimulated to have an immune response against the diseased cells.Once the host's own immune system is stimulated to have such a response,other diseased cells that are not treated by the activatablepharmaceutical agent may be recognized and be destroyed by the host'sown immune system. Such autovaccine effects may be obtained, forexample, in treatments using psoralen and UV-A.

In another aspect, the present invention also provides methods forproducing an autovaccine, including: (1) providing a population oftargeted cells; (2) treating the cells ex vivo with a psoralen or aderivative thereof; (3) activating the psoralen with a UV-A source toinduce apoptosis in the targeted cells; and (4) returning the apopticcells back to the host to induce an autovaccine effect against thetargeted cell, wherein the apoptic cells cause an autovaccine effect.

A further embodiment is the use of the present invention for thetreatment of skin cancer. In this example, a photoactivatable agent,preferably psoralen, is given to the patient, and is delivered to theskin lesion via the blood supply. An activation source having limitedpenetration ability (such as UV or IR) is shined directly on the skin—inthe case of psoralen, it would be a UV light, or an IR source. With theuse of an IR source, the irradiation would penetrate deeper and generateUV via two single photon events with psoralen.

In a further embodiment, methods according to this aspect of the presentinvention further include a step of separating the components of apopticcells into fractions and testing each fraction for autovaccine effect ina host. The components thus isolated and identified may then serve as aneffective autovaccine to stimulate the host's immune system to suppressgrowth of the targeted cells.

The present invention methods can be used alone or in combination withother therapies for treatment of cell proliferation disorders.Additionally, the present invention methods can be used, if desired, inconjunction with recent advances in chronomedicine, such as thatdetailed in Giacchetti et al, Journal of Clinical Oncology, Vol 24, No22 Aug. 1, 2006: pp. 3562-3569. In chronomedicine it has been found thatcells suffering from certain types of disorders, such as cancer, respondbetter at certain times of the day than at others. Thus, chronomedicinecould be used in conjunction with the present methods in order toaugment the effect of the treatments of the present invention.

In another aspect, the present invention further provides systems andkits for practicing the above described methods.

In one embodiment, a system in accordance with the present invention mayinclude: (1) an initiation energy source; (2) one or more energymodulation agents; and (3) one or more activatable pharmaceuticalagents.

In another embodiment, a system in accordance with the present inventionmay include an initiation energy source and one or more activatablepharmaceutical agents.

FIG. 3G illustrates a system according to one exemplary embodiment ofthe present invention. Referring to FIG. 3G, an exemplary systemaccording to one embodiment of the present invention may have aninitiation energy source 1 directed at the subject 4. An activatablepharmaceutical agent 2 and an energy modulation agent 3 are administeredto the subject 4. The initiation energy source may additionally becontrolled by a computer system 5 that is capable of directing thedelivery of the initiation energy.

In preferred embodiments, the initiation energy source may be a linearaccelerator equipped with image guided computer-control capability todeliver a precisely calibrated beam of radiation to a pre-selectedcoordinate. One example of such linear accelerators is the SmartBeam™IMRT (intensity modulated radiation therapy) system from Varian medicalsystems (Varian Medical Systems, Inc., Palo Alto, Calif.).

In other embodiments, endoscopic or laproscopic devices equipped withappropriate initiation energy emitter may be used as the initiationenergy source. In such systems, the initiation energy may be navigatedand positioned at the pre-selected coordinate to deliver the desiredamount of initiation energy to the site.

In further embodiments, dose calculation and robotic manipulationdevices may also be included in the system.

In yet another embodiment, there is also provided a computer implementedsystem for designing and selecting suitable combinations of initiationenergy source, energy transfer agent, and activatable pharmaceuticalagent, comprising:

a central processing unit (CPU) having a storage medium on which isprovided:

-   -   a database of excitable compounds;    -   a first computation module for identifying and designing an        excitable compound that is capable of binding with a target        cellular structure or component; and    -   a second computation module predicting the resonance absorption        energy of the excitable compound,

wherein the system, upon selection of a target cellular structure orcomponent, computes an excitable compound that is capable of bindingwith the target structure followed by a computation to predict theresonance absorption energy of the excitable compound.

FIG. 4 illustrates an exemplary computer implemented system according tothis embodiment of the present invention. Referring to FIG. 4, anexemplary computer-implemented system according to one embodiment of thepresent invention may have a central processing unit (CPU) connected toa memory unit, configured such that the CPU is capable of processinguser inputs and selecting a combination of initiation source,activatable pharmaceutical agent, and energy transfer agent based on anenergy spectrum comparison for use in a method of the present invention.

FIG. 5 illustrates a computer system 1201 for implementing variousembodiments of the present invention. The computer system 1201 may beused as the controller 55 to perform any or all of the functions of theCPU described above. The computer system 1201 includes a bus 1202 orother communication mechanism for communicating information, and aprocessor 1203 coupled with the bus 1202 for processing the information.The computer system 1201 also includes a main memory 1204, such as arandom access memory (RAM) or other dynamic storage device (e.g.,dynamic RAM (DRAM), static RAM (SRAM), and synchronous DRAM (SDRAM)),coupled to the bus 1202 for storing information and instructions to beexecuted by processor 1203. In addition, the main memory 1204 may beused for storing temporary variables or other intermediate informationduring the execution of instructions by the processor 1203. The computersystem 1201 further includes a read only memory (ROM) 1205 or otherstatic storage device (e.g., programmable ROM (PROM), erasable PROM(EPROM), and electrically erasable PROM (EEPROM)) coupled to the bus1202 for storing static information and instructions for the processor1203.

The computer system 1201 also includes a disk controller 1206 coupled tothe bus 1202 to control one or more storage devices for storinginformation and instructions, such as a magnetic hard disk 1207, and aremovable media drive 1208 (e.g., floppy disk drive, read-only compactdisc drive, read/write compact disc drive, compact disc jukebox, tapedrive, and removable magneto-optical drive). The storage devices may beadded to the computer system 1201 using an appropriate device interface(e.g., small computer system interface (SCSI), integrated deviceelectronics (IDE), enhanced-IDE (E-IDE), direct memory access (DMA), orultra-DMA).

The computer system 1201 may also include special purpose logic devices(e.g., application specific integrated circuits (ASICs)) or configurablelogic devices (e.g., simple programmable logic devices (SPLDs), complexprogrammable logic devices (CPLDs), and field programmable gate arrays(FPGAs)).

The computer system 1201 may also include a display controller 1209coupled to the bus 1202 to control a display 1210, such as a cathode raytube (CRT), for displaying information to a computer user. The computersystem includes input devices, such as a keyboard 1211 and a pointingdevice 1212, for interacting with a computer user and providinginformation to the processor 1203. The pointing device 1212, forexample, may be a mouse, a trackball, or a pointing stick forcommunicating direction information and command selections to theprocessor 1203 and for controlling cursor movement on the display 1210.In addition, a printer may provide printed listings of data storedand/or generated by the computer system 1201.

The computer system 1201 performs a portion or all of the processingsteps of the invention (such as for example those described in relationto FIG. 5) in response to the processor 1203 executing one or moresequences of one or more instructions contained in a memory, such as themain memory 1204. Such instructions may be read into the main memory1204 from another computer readable medium, such as a hard disk 1207 ora removable media drive 1208. One or more processors in amulti-processing arrangement may also be employed to execute thesequences of instructions contained in main memory 1204. In alternativeembodiments, hard-wired circuitry may be used in place of or incombination with software instructions. Thus, embodiments are notlimited to any specific combination of hardware circuitry and software.

As stated above, the computer system 1201 includes at least one computerreadable medium or memory for holding instructions programmed accordingto the teachings of the invention and for containing data structures,tables, records, or other data described herein. Examples of computerreadable media are compact discs, hard disks, floppy disks, tape,magneto-optical disks, PROMs (EPROM, EEPROM, flash EPROM), DRAM, SRAM,SDRAM, or any other magnetic medium, compact discs (e.g., CD-ROM), orany other optical medium, punch cards, paper tape, or other physicalmedium with patterns of holes, a carrier wave (described below), or anyother medium from which a computer can read.

Stored on any one or on a combination of computer readable media, thepresent invention includes software for controlling the computer system1201, for driving a device or devices for implementing the invention,and for enabling the computer system 1201 to interact with a human user(e.g., print production personnel). Such software may include, but isnot limited to, device drivers, operating systems, development tools,and applications software. Such computer readable media further includesthe computer program product of the present invention for performing allor a portion (if processing is distributed) of the processing performedin implementing the invention.

The computer code devices of the present invention may be anyinterpretable or executable code mechanism, including but not limited toscripts, interpretable programs, dynamic link libraries (DLLs), Javaclasses, and complete executable programs. Moreover, parts of theprocessing of the present invention may be distributed for betterperformance, reliability, and/or cost.

The term “computer readable medium” as used herein refers to any mediumthat participates in providing instructions to the processor 1203 forexecution. A computer readable medium may take many forms, including butnot limited to, non-volatile media, volatile media, and transmissionmedia. Non-volatile media includes, for example, optical, magneticdisks, and magneto-optical disks, such as the hard disk 1207 or theremovable media drive 1208. Volatile media includes dynamic memory, suchas the main memory 1204. Transmission media includes coaxial cables,copper wire and fiber optics, including the wires that make up the bus1202. Transmission media also may also take the form of acoustic orlight waves, such as those generated during radio wave and infrared datacommunications.

Various forms of computer readable media may be involved in carrying outone or more sequences of one or more instructions to processor 1203 forexecution. For example, the instructions may initially be carried on amagnetic disk of a remote computer. The remote computer can load theinstructions for implementing all or a portion of the present inventionremotely into a dynamic memory and send the instructions over atelephone line using a modem. A modem local to the computer system 1201may receive the data on the telephone line and use an infraredtransmitter to convert the data to an infrared signal. An infrareddetector coupled to the bus 1202 can receive the data carried in theinfrared signal and place the data on the bus 1202. The bus 1202 carriesthe data to the main memory 1204, from which the processor 1203retrieves and executes the instructions. The instructions received bythe main memory 1204 may optionally be stored on storage device 1207 or1208 either before or after execution by processor 1203.

The computer system 1201 also includes a communication interface 1213coupled to the bus 1202. The communication interface 1213 provides atwo-way data communication coupling to a network link 1214 that isconnected to, for example, a local area network (LAN) 1215, or toanother communications network 1216 such as the Internet. For example,the communication interface 1213 may be a network interface card toattach to any packet switched LAN. As another example, the communicationinterface 1213 may be an asymmetrical digital subscriber line (ADSL)card, an integrated services digital network (ISDN) card or a modem toprovide a data communication connection to a corresponding type ofcommunications line. Wireless links may also be implemented. In any suchimplementation, the communication interface 1213 sends and receiveselectrical, electromagnetic or optical signals that carry digital datastreams representing various types of information.

The network link 1214 typically provides data communication through oneor more networks to other data devices. For example, the network link1214 may provide a connection to another computer through a localnetwork 1215 (e.g., a LAN) or through equipment operated by a serviceprovider, which provides communication services through a communicationsnetwork 1216. The local network 1214 and the communications network 1216use, for example, electrical, electromagnetic, or optical signals thatcarry digital data streams, and the associated physical layer (e.g., CAT5 cable, coaxial cable, optical fiber, etc). The signals through thevarious networks and the signals on the network link 1214 and throughthe communication interface 1213, which carry the digital data to andfrom the computer system 1201 maybe implemented in baseband signals, orcarrier wave based signals. The baseband signals convey the digital dataas unmodulated electrical pulses that are descriptive of a stream ofdigital data bits, where the term “bits” is to be construed broadly tomean symbol, where each symbol conveys at least one or more informationbits. The digital data may also be used to modulate a carrier wave, suchas with amplitude, phase and/or frequency shift keyed signals that arepropagated over a conductive media, or transmitted as electromagneticwaves through a propagation medium. Thus, the digital data may be sentas unmodulated baseband data through a “wired” communication channeland/or sent within a predetermined frequency band, different thanbaseband, by modulating a carrier wave. The computer system 1201 cantransmit and receive data, including program code, through thenetwork(s) 1215 and 1216, the network link 1214, and the communicationinterface 1213. Moreover, the network link 1214 may provide a connectionthrough a LAN 1215 to a mobile device 1217 such as a personal digitalassistant (PDA) laptop computer, or cellular telephone.

The exemplary energy spectrum previously noted in FIG. 1 may also beused in this computer-implemented system.

The reagents and chemicals useful for methods and systems of the presentinvention may be packaged in kits to facilitate application of thepresent invention. In one exemplary embodiment, a kit including apsoralen, and fractionating containers for easy fractionation andisolation of autovaccines is contemplated. A further embodiment of kitwould comprise at least one activatable pharmaceutical agent capable ofcausing a predetermined cellular change, at least one energy modulationagent capable of activating the at least one activatable agent whenenergized, and containers suitable for storing the agents in stableform, and preferably further comprising instructions for administeringthe at least one activatable pharmaceutical agent and at least oneenergy modulation agent to a subject, and for applying an initiationenergy from an initiation energy source to activate the activatablepharmaceutical agent. The instructions could be in any desired form,including but not limited to, printed on a kit insert, printed on one ormore containers, as well as electronically stored instructions providedon an electronic storage medium, such as a computer readable storagemedium. Also optionally included is a software package on a computerreadable storage medium that permits the user to integrate theinformation and calculate a control dose, to calculate and controlintensity of the irradiation source.

Having generally described this invention, a further understanding canbe obtained by reference to certain specific examples which are providedherein for purposes of illustration only and are not intended to belimiting unless otherwise specified.

EXAMPLES Example 1

In a first example, Vitamin B12 is used as a stimulating energy sourcefor a photoactive agent overlapping its emission wavelength usingdipole-dipole resonance energy transfer.

Excitation Max. Emission Max. Endogenous Fluorophore (nm) (nm) VitaminB₁₂ 275 305

Vitamin B12 has an excitation maximum at about 275 nm and an emissionmaximum at 305 nm, as shown above and in Table 2. Table 4 shows UV andlight emission from gamma ray sources. In this example, ¹¹³Sn and/or¹³⁷Cs are chelated with the Vitamin B12. The Vitamin B12 preferentiallyis absorbed by tumor cells. Thus, it is in close proximity and capableof activating 8-MOP, which is administered in advance as thephotoactivation molecules. The emission band of Vitamin B12 overlaps theexcitation band of 8-MOP; therefore, photo and resonance energy transferoccurs, when Vitamin B12 is in close proximity to 8-MOP. 8-MOP isactivated and binds to DNA of the tumor cells inducing an auto vaccineeffect in vivo.

Example 2

In this example, gold nanoparticles are chelated with the Vitamin B12complex. A suitable light source is used to stimulate the goldnanoparticles or Vitamin B12 may be chelated with one of the UV emitterslisted in Table 4 in addition to the gold nanoparticles. The tumor cellspreferentially absorb the Vitamin B12 complexes, such that the activatedgold nanoparticles are within 50 nanometers of 8-MOP and/or otherphotoactivatable molecules previously administered. Therefore, resonanceenergy transfer activates the photoactivatable molecules, such as 8-MOP,and the activated 8-MOP binds to DNA in tumor cells indusing apoptosisand autovaccine effects.

In a further example, the nanoparticles of gold are clusters of 5 goldatoms encapsulated by poly-amidoamine dendrimers. Thus, the goldnanoparticles emit UV in the correct band for activating 8-MOP and otherUV-activatable agents capable of exhibiting photophoresis and/orphotodynamic effects.

Cells undergoing rapid proliferation have been shown to have increaseduptake of thymidine and methionine. (See, for example, M. E. vanEijkeren et al., Acta Oncologica, 31, 539 (1992); K. Kobota et al., J.Nucl. Med., 32, 2118 (1991) and K. Higashi et al., J. Nucl. Med., 34,773(1993)). Since methylcobalamin is directly involved with methioninesynthesis and indirectly involved in the synthesis of thymidylate andDNA, it is not surprising that methylcobalamin as well asCobalt-57-cyanocobalamin have also been shown to have increased uptakein rapidly dividing tissue (for example, see, B. A. Cooper et al.,Nature, 191, 393 (1961); H. Flodh, Acta Radiol. Suppl., 284, 55 (1968);L. Bloomquist et al., Experientia, 25, 294 (1969)). Additionally, upregulation in the number of transcobalamin I1 receptors has beendemonstrated in several malignant cell lines during their acceleratedthymidine incorporation and DNA synthesis (see, J. Lindemans et al.,Exp. Cell. Res., 184, 449 (1989); T. Amagasaki et al., Blood, 26, 138(1990) and J. A. Begly et al., J. Cell Physiol. 156, 43 (1993). VitaminB12 is water soluble, has no known toxicity, and in excess is excretedby gloinerular filtration. In addition, the uptake of vitamin B12 couldpotentially be manipulated by the administration of nitrous oxide andother pharmacological agents (D. Swanson et al., Pharmaceuticals inMedical Imaging, MacMillan Pub. Co., NY (1990) at pages 621 628).

A preferred embodiment of the present invention uses a psoralen compoundas the activatable pharmaceutical agent (most preferably 8-MOP or AMT),nanoparticles of gold having clusters of 5 gold atoms encapsulated bypoly-amidoamine dendrimers as the energy modulation agent, x-rays as theinitiation energy source, UV-A as the resultant energy emitted by theenergy modulation agent, which upon activation of the psoralen compoundresults in apoptosis in the target cells.

Waste Water Detoxification

Photocatalysis has also been used as tertiary treatment for wastewaterto comply with the regulatory discharge limits and to oxidize persistentcompounds that have not been oxidized in the biological treatment.Photocatalysis has being applied to the elimination of severalpollutants (e.g., alkanes, alkenes, phenols, aromatics, pesticides) withgreat success. In many cases, total mineralization of the organiccompounds has been observed. Several photocatalysts, such as CdS, Fe₂O₃,ZnO, WO₃, and ZnS, have been studied, but the best results have beenachieved with TiO₂P₂₅. These photocatalyst are usable for the inventiondescribed here.

The wastewaters of an oil refinery are the waters resulting from washingthe equipment used in the process, undesirable wastes, and sanitarysewage. These effluents have high oil and grease contents, besides otherorganic compounds in solution. These pollutants form a residual chemicaloxygen demand (COD) that may pose serious toxic hazards to theenvironment.

It is known that photocatalysis can be used for waste water reductionremediation. U.S. Pat. No. 5,118,422 (the entire contents of which areincorporated herein by reference) to Cooper et al. describe anultraviolet driven photocatalytic post-treatment technique for purifyinga water feedstock containing an oxidizable contaminant compound. In thiswork, the water feedstock was mixed with photocatalytic semiconductorparticles (e.g., TiO₂, ZnO, CdS, CdSe, SnO₂, SrTiO₃, WO₃, Fe₂O₃, andTa₂O₅ particles) having a particle size in the range of about 0.01 toabout 1.0 micron and in an amount of between about 0.01% and about 0.2%by weight of the water. The water including the semiconductor mixture isexposed to band-gap photons for a time sufficient to effect an oxidationof the oxidizable contaminant to purify the water. Crossflow membranefiltration was used to separate the purified water from thesemiconductor particles. Cooper et al. show that the organic impuritycarbon content of simulated reclamation waters at nominal 40 PPM levelwere reduced to parts per billion using a recirculation batch reactor.

Cooper et al. identified that one important aspect of the photocatalyticprocess is the adsorption of the organic molecules onto the extremelylarge surface area presented by the finely divided powders dispersed inthe water. Cooper et al. further indicated that, in photoelectrochemicalapplications, advantage is taken of the fact that the solid phase (ametal oxide semiconductor) is also photo-active and that the generatedcharge carriers are directly involved in the organic oxidation. Theadsorption of the band-gap photon by the semiconductor particle resultsin the formation of an electron (e⁻)/hole (h⁺) pair. Cooper et al.explain that the electrons generated in the conduction band react withsolution oxygen forming the dioxygen anion (O²⁻) species whichsubsequently undergo further reactions resulting in the production ofthe powerfully oxidizing hydroxyl radical species, OH. These powerfuloxidants are known to oxidize organic compounds by themselves.Additionally, Cooper et al. explain that the strongly oxidizing holesgenerated in the valence band have sufficient energy to oxidize allorganic bonds.

In the reactor of Cooper et al., turbulence is necessary in order toensure that the waste water contaminants and the photocatalytic titaniaparticles are exposed to the UV light. Cooper et al. explain that themost basic considerations of photocatalyst light adsorption and itsrelationship to convective mixing. For a 0.1 wt % photocatalyst loading,experiments have shown that 90% of the light is absorbed within 0.08 cm.This is primarily due to the large UV absorption coefficient of thephotocatalyst and therefore, most of the photoelectrochemistry occurswithin this illuminated region. By operating the reactor of Cooper etal. with a Reynolds number (Re) of 4000, a significant portion of thephotoactive region is ensured of being within the well mixed turbulentzone.

Santos et al. have reported in “Photocatalysis as a tertiary treatmentfor petroleum refinery wastewaters” published in Braz. J. Chem. Eng.vol. 23, No. 4, 2006 (the entire contents of which are incorporatedherein by reference), photocatalysis for tertiary treatment forpetroleum refinery wastewaters which satisfactorily reduced the amountof pollutants to the level of the regulatory discharge limits andoxidized persistent compounds that had not been oxidized in thebiological treatment. The treatment sequence used by the refinery(REDUC/PETROBRAS, a Brazilian oil refinery) is oil/water separationfollowed by a biological treatment. Although the process efficiency interms of biological oxygen demand (BOD) removal is high, a residual andpersistent COD and a phenol content remains. The refining capacity ofthe refinery is 41,000 m³/day, generating 1,100 m³/h of wastewater,which are discharged directly into the Guanabara Bay (Rio de Janeiro).Treating the residual and persistent COD remains a priority.

Santos et al. conducted a first set of experiments carried out in anopen 250 mL reactor containing 60 mL of wastewater. In the second set ofexperiments, a Pyrex® annular reactor containing 550 mL of wastewaterwas used (De Paoli and Rodrigues, 1978), as shown in FIG. 1. Thereaction mixtures inside the reactors were maintained in suspension bymagnetic stirring. In all experiments, air was continuously bubbledthrough the suspensions. A 250 W Phillips HPL-N medium pressure mercuryvapor lamp (with its outer bulb removed) was used as the UV-light source(radiant flux of 108 J·m⁻²·s⁻¹ at 8>254 nm). In one set of experiments,the lamp was positioned above the surface of the liquid at a fixedheight (12 cm). In the second set, the lamp was inserted into the well.All experiments by Santos et al. were performed at 25+1° C. The catalystconcentration ranged from 0.5 to 5.5 g L⁻¹ and the initial pH rangedfrom 3.5 to 9.

In the invention described herein, luminescing particles or other energymodulation agents would be placed inside quartz or glass fixtures withinthe waste water or would be placed on silica encapsulated structureswithin the waste water which, like the photocatalytic TiO₂, could beentrained in the waste water during the irradiation.

Upon irradiation with x-rays (or other penetrating radiation) throughfor example a plastic or aluminum container, activation of theluminescing particles (i.e., energy modulation agents) would generate UVlight in nearby presence of the photocatalytic agent. In other words forthe invention described herein, the luminescent particles or otherenergy modulation agents are mixed along with the photocatalyticsemiconductor particles in the waste water fluid stream, and theexterior activation energy source penetrates the container (e.g., aplastic or aluminum container) and irradiates the bulk of the wastewater, producing UV light throughout the waste water which in turndrives the photocatalytic reactions.

As such, the invention described herein offers a number to advantagesover that described above, including the elimination of expensiveholding tanks for the waste water, the avoidance of having to pump thewastewater at higher pressures or flowrates to produce sufficientturbulence, and the generation of UV light throughout the wastewater tothereby provide faster bulk processing of the waste water.

Photostimulation

Photostimulation is a field in which light is applied to in order toalter or change a physical property. For example, there has been anincreased focus on the use of biodegradable polymers in consumer andbiomedical fields. Polylactic acid (PLA) plastics andpolyhydroxyalkanoates (PHA) plastics have been playing a vital role infulfilling the objectives. But their relatively hydrophobic surfaceslimit their use in various applications. Hence, there is a need tosurface modify these film surfaces. Due to the lack of any modifiableside chain groups, workers have used a sequential two step photograftingtechnique for the surface modification of these biopolymers. In stepone, benzophenone was photografted on the film surface and in step two,hydrophilic monomers like acrylic acid and acrylamide werephotopolymerized from the film surfaces.

Workers have found that UV irradiation could realize an effective graftcopolymerization. UV-assisted photografting in ethanol has been used togrow hydrophilic polymers (e.g., poly(acrylic acid) and polyacrylamide)from the surfaces of PLA, PHA, and PLA/PHA blend films. In that work, afunctional polyurethane (PU) surface was prepared by photo-graftingN,N-dimethylaminoethyl methacrylate (DMAEM) onto the membrane surface.Grafting copolymerization was conducted by the combined use of thephoto-oxidation and irradiation grafting. PU membrane was photo-oxidizedto introduce the hydroperoxide groups onto the surface, then themembrane previously immersed in monomer solution was irradiated by UVlight. Results have shown prior to the invention that UV irradiation canrealize graft copolymerization effectively.

In the invention described herein, these processes are expedited by theinclusion of luminescing particles or other energy modulation agents indispersion in the fluid medium being used for photostimulation.

Upon irradiation with x-rays (or other penetrating radiation) throughfor example a plastic or aluminum container, activation of theluminescing particles (i.e., energy modulation agents) would generate UVlight throughout the volume of the medium (eliminating any shadowingeffects) and permitting batch or bulk type processing to occur inparallel throughout the container.

In other examples, the interior generation of light inside a bulk mediummay serve to stimulate a chemical or biological process either by directinteraction of the light with activatable agents in the medium or theindirect generation of heat which the invention described here by way ofdispersed energy modulation agents would provide a controlled anduniform way to heat a vat of material in a biological or chemicalprocess.

Photodeactivation

In many industrial processes, especially food and beverage industries,yeasts are used to produce changes in a medium such as the conversion ofsugars in the raw product. One particularly prominent example is in thewine industry. Stopping the wine from fermenting any further wouldpreserve the current level of sweetness. Likewise, allowing the wine tocontinue fermenting further would only make the wine less sweet witheach passing day. Eventually the wine would become completely dry atwhich time the fermentation would stop on its own. This is becauseduring the fermentation process yeast turns the sugar into alcohol.

Wanting to stop a fermentation is all good in and of itself. Butunfortunately, there is really no practical way to successfully stop afermentation dead in its tracks. Additives such as sulphite and sorbatecan be added to stabilize a fermented product and stop additionalfermentation. Many winemakers will turn to sulfites such as that foundin Sodium Bisulfite or Campden tablets for the answer. But, these twoitems are not capable of reliably killing enough of the yeast toguarantee a complete stop of the activity—at least not at normal dosesthat leave the wine still drinkable.

Once the bulk of the sulfites from either of these ingredients dissipatefrom the wine into the air—as sulfites do—there is a very strong chancethat the remaining few live yeast cells will start multiplying andfermenting again if given enough time. This usually happens at a mostinconvenient time, like after the wine has been bottled and stowed away.

Potassium sorbate is another ingredient that many winemakers considerwhen trying to stop a wine from fermenting any further. There is a lotof misunderstanding surrounding this product. It is typically called forby home wine making books when sweetening a wine. This is a situationwhere the fermentation has already completed and is ready for bottling.One adds the potassium sorbate along with the sugar that is added forsweetening.

The potassium sorbate stops the yeast from fermenting the newly addedsugar. So, many winemakers assume potassium sorbate can stop an activefermentation as well, but, potassium sorbate does not kill the yeast atall, but rather it makes the yeast sterile. In other words, it impairsthe yeast's ability to reproduce itself. But, it does not hinder theyeast's ability to ferment sugar into alcohol.

Ultraviolet light is known to destroy yeast cultures, but has restrictedapplications due to the inability of UV light to penetrate throughoutthe fluid medium. While heat can be used to destroy the yeast activity,cooking of the product may be premature or may produce undesirablechanges in the consistency and taste. For liquid or fluid food products,the same techniques described above for liquid pasteurization could beused for the invention described here. For non-liquid products, energymodulation agents with little and preferably no toxicity (e.g. Fe oxidesor titanium oxides) could be added. Here, the concentration of theseadditives would likely be limited by any unexpected changes in taste.

Photoactivated Cross-Linking and Curing of Polymers

In this application, luminescing particles (or energy modulation agents)are provided and distributed into an uncured polymer based medium forthe activation of photosensitive agents in the medium to promotecross-linking and curing of the polymer based medium.

As noted above, for adhesive and surface coating applications, lightactivated processing is limited due to the penetration depth of UV lightinto the processed medium. In light activated adhesive and surfacecoating processing, the primary limitation is that the material to becured must see the light—both in type (wavelength or spectraldistribution) and intensity. This limitation has meant that one mediumtypically has to transmit the appropriate light. In adhesive and surfacecoating applications, any “shaded” area will require a secondary curemechanism, increasing cure time over the non-shaded areas and furtherdelaying cure time due to the existent of a sealed skin through whichsubsequent curing must proceed.

Conventionally, moisture-curing mechanisms, heat-curing mechanisms, andphoto-initiated curing mechanisms are used to initiate cure, i.e.,cross-linking, of reactive compositions, such as reactive silicones,polymers, and adhesives. These mechanisms are based on eithercondensation reactions, whereby moisture hydrolyzes certain groups, oraddition reactions that can be initiated by a form of energy, such aselectromagnetic radiation or heat.

The invention described herein can use any of the following lightactivated curing polymers as well as others known in the art to whichthe luminescing particles (or energy modulation agents) are added.

For example, one suitable light activated polymer compound includes UVcuring silicones having methacrylate functional groups. U.S. Pat. No.4,675,346 to Lin, the disclosure of which is hereby expresslyincorporated herein by reference, is directed to UV curable siliconecompositions including at least 50% of a specific type of siliconeresin, at least 10% of a fumed silica filler and a photoinitiator, andcured compositions thereof. Other known UV curing silicone compositionssuitable for the invention include organopolysiloxane containing a(meth)acrylate functional group, a photosensitizer, and a solvent, whichcures to a hard film. Other known UV curing silicone compositionssuitable for the invention include compositions of an organopolysiloxanehaving an average of at least one acryloxy and/or methacryloxy group permolecule; a low molecular weight polyacrylyl crosslinking agent; and aphotosensitizer.

Loctite Corporation has designed and developed UV and UV/moisture dualcurable silicone compositions, which also demonstrate high resistance toflammability and combustibility, where the flame-retardant component isa combination of hydrated alumina and a member selected from the groupconsisting of organo ligand complexes of transition metals,organosiloxane ligand complexes of transition metals, and combinationsthereof. See U.S. Pat. Nos. 6,281,261 and 6,323,253 to Bennington. Theseformulations are also suitable for the invention.

Other known UV photoactivatable silicones include siliconesfunctionalized with for example carboxylate, maleate, cinnamate andcombinations thereof. These formulations are also suitable for theinvention. Other known UV photoactivatable silicones suitable for theinvention include benzoin ethers (“UV free radical generator”) and afree-radical polymerizable functional silicone polymers, as described inU.S. Pat. No. 6,051,625 whose content is incorporated herein byreference in its entirety. The UV free radical generator (i.e., thebenzoin ether) is contained at from 0.001 to 10 wt % based on the totalweight of the curable composition. Free radicals produced by irradiatingthe composition function as initiators of the polymerization reaction,and the free radical generator can be added in a catalytic quantityrelative to the polymerizable functionality in the subject composition.Further included in these silicone resins can be silicon-bonded divalentoxygen atom compounds which can form a siloxane bond while the remainingoxygen in each case can be bonded to another silicon to form a siloxanebond, or can be bonded to methyl or ethyl to form an alkoxy group, orcan be bonded to hydrogen to form silanol. Such compounds can includetrimethylsilyl, dimethylsilyl, phenyldimethylsilyl, vinyldimethylsilyl,trifluoropropyldimethylsilyl, (4-vinylphenyl)dimethylsilyl,(vinylbenzyl)dimethylsilyl, and (vinylphenethyl)dimethylsilyl.

The photoinitiator component of the invention is not limited to thosefree radical generators given above, but may be any photoinitiator knownin the art, including the afore-mentioned benzoin and substitutedbenzoins (such as alkyl ester substituted benzoins), Michler's ketone,dialkoxyacetophenones, such as diethoxyacetophenone (“DEAP”),benzophenone and substituted benzophenones, acetophenone and substitutedacetophenones, and xanthone and substituted xanthones. Other desirablephotoinitiators include DEAP, benzoin methyl ether, benzoin ethyl ether,benzoin isopropyl ether, diethoxyxanthone, chloro-thio-xanthone,azo-bisisobutyronitrile, N-methyl diethanolaminebenzophenone, andmixtures thereof. Visible light initiators include camphoquinone,peroxyester initiators and non-fluorene-carboxylic acid peroxyesters.

Commercially available examples of photoinitiators suitable for theinvention include those from Vantico, Inc., Brewster, N.Y. under theIRGACURE and DAROCUR tradenames, specifically IRGACURE 184(1-hydroxycyclohexyl phenyl ketone), 907(2-methyl-1-[4-(methylthio)phenyl]-2-morpholino propan-1-one), 369(2-benzyl-2-N,N-dimethylamino-1-(4-morpholinophenyl)-1-butanone), 500(the combination of 1-hydroxy cyclohexyl phenyl ketone andbenzophenone), 651 (2,2-dimethoxy-2-phenyl acetophenone), 1700 (thecombination of bis(2,6-dimethoxybenzoyl-2,4,4-trimethyl pentyl)phosphineoxide and 2-hydroxy-2-methyl-1-phenyl-propan-1-one), and 819[bis(2,4,6-trimethyl benzoyl)phenyl phosphine oxide] and DAROCUR 1173(2-hydroxy-2-methyl-1-phenyl-1-propane) and 4265 (the combination of2,4,6-trimethylbenzoyldiphenyl-phosphine oxide and2-hydroxy-2-methyl-1-phenyl-propan-1-one); and IRGACURE 784DC(bis(.eta..sup.5-2,4-cyclopentadien-1-yl)-bis[2,6-difluoro-3-(1H-pyrrol-1-yl)phenyl]titanium).

Generally, the amount of photoinitiator (or free radical generators)should be in the range of about 0.1% to about 10% by weight, such asabout 2 to about 6% by weight. The free radical generator concentrationfor benzoin ether is generally from 0.01 to 5% based on the total weightof the curable composition.

A moisture cure catalyst can also be included in an amount effective tocure the composition. For example, from about 0.1 to about 5% by weight,such as about 0.25 to about 2.5% by weight, of the moisture curecatalyst can be used in the invention to facilitate the cure processbeyond that of photo-activated curing. Examples of such catalystsinclude organic compounds of titanium, tin, zirconium and combinationsthereof. Tetraisopropoxytitanate and tetrabutoxytitanate are suitable asmoisture cure catalyst. See also U.S. Pat. No. 4,111,890, the disclosureof which is expressly incorporated herein by reference.

Included in the conventional silicone composition (and other inorganicand organic adhesive polymers) suitable for the invention are variousinorganic fillers. For example, hollow microspheres supplied by Kishunder the trade name Q-CEL are free flowing powders, white in color.Generally, these borosilicate hollow microspheres are promoted asextenders in reactive resin systems, ordinarily to replace heavyfillers, such as calcium carbonate, thereby lowering the weight ofcomposite materials formed therewith. Q-CEL 5019 hollow microspheres areconstructed of a borosilicate, with a liquid displacement density of0.19 g/cm², a mean particle size of 70 microns, and a particle sizerange of 10-150 urn. Other Q-CEL products are shown below in tabularform. Another commercially available hollow glass microsphere is sold byKish under the trade name SPHERICEL. SPHEREICEL 110P8 has a meanparticle size of about 11.7 microns, and a crush strength of greaterthan 10,000 psi. Yet other commercially available hollow glassmicrosphere are sold by the Schundler Company, Metuchen, N.J. under thePERLITE tradename, Whitehouse Scientific Ltd., Chester, UK and 3M,Minneapolis, Minn. under the SCOTCHLITE tradename.

In general, these inorganic filler components (and others such as fumedsilica) add structural properties to the cured composition, as well asconfers flowability properties to the composition in the uncured stateand increase the transmissivity for the UV cure radiation. When present,the fumed silica can be used at a level of up to about 50 weightpercent, with a range of about 4 to at least about 10 weight percent,being desirable. While the precise level of silica may vary depending onthe characteristics of the particular silica and the desired propertiesof the composition and the reaction product thereof, care should beexercised by those persons of ordinary skill in the art to allow for anappropriate level of transmissivity of the inventive compositions topermit a UV cure to occur.

Desirable hydrophobic silicas include hexamethyldisilazane-treatedsilicas, such as those commercially available from Wacker-Chemie,Adrian, Mich. under the trade designation HDK-2000. Others includepolydimethylsiloxane-treated silicas, such as those commerciallyavailable from Cabot Corporation under the trade designation CAB-O-SILN70-TS, or Degussa Corporation under the trade designation AEROSIL R202.Still other silicas include trialkoxyalkyl silane-treated silicas, suchas the trimethoxyoctyl silane-treated silica commercially available fromDegussa under the trade designation AEROSIL R805; and 3-dimethyldichlorosilane-treated silicas commercially available from Degussa underthe trade designation R972, R974 and R976.

While these inorganic fillers have extended the use of conventional UVcured silicone systems to permit the curing of materials beyond a skindepth of UV penetration, these inorganic fillers alone do not overcomeshadowing effects and suffer from UV scattering which effectively makesfor a smaller penetration depth. In the invention described herein, theinclusion of these inorganic fillers along with luminescing particlesprovide a mechanism by which uniform light activated cures can occurdeep inside of the body of adhesive-solidified assemblies in regionsthat would normally be shadowed or not with the reach of external UV orother light sources.

Accordingly, in this example of the invention described herein,conventional silicone and polymeric adhesive or release or coatingcompositions are prepared using conventional mixing, heating, andincubation techniques. Included in these conventional compositions areluminescing particles. These luminescing particle containingcompositions can then be applied to surfaces of objects to be fixedtogether or to surfaces where a hard coating is desired or cast in acurable form for the production of molded objects. The luminescingparticles in these compositions upon activation will produce radiantlight for photoactivated cure of the luminescing particle containingpolymer composition. The density of luminescing particles in thesecompositions will depend on the “light transparency” of the luminescingparticle containing composition. Where these compositions contain asignificant amount of the inorganic filler as discussed above, theconcentration of luminescing particles can be reduced for example ascompared to a composition with a black color pigment where the lighttransparency will be significantly reduced.

One advantage of the invention described here as seen from this exampleis that now color pigments can be included in the light curable resinswithout significant compromise in the cured product performance. Thesecolor pigments may include one or more colored pigments well known tothose of ordinary skill in the art. Such pigments are generally metaloxides and include, but are not limited to, titanium dioxide, ironoxides, organic complexes, mica, talc and quartz. One pigment may beused, or a combination of two or more pigments may be utilized.Different colors can be obtained by choosing proper pigments andcombining them in a similar fashion as set forth in the followingexamples with the necessary adjustments, common in the paint industry,being made. Accordingly, in one embodiment of the invention, these colorpigments including carbon black may also be included as an opticallyopaque materials to limit the propagation of internally generated lightfrom the point of generation.

U.S. Pat. No. 7,294,656 to Bach et al., the entire disclosure of whichis incorporated herein by reference, describes a non-aqueous compositioncurable by UV radiation broadly containing a mixture of two UV curableurethane acrylates that have several advantages over conventionalradiation-curable compositions. The Bache et al. compositions can becured in a relatively short time using UV-C (200-280 nm), UV-B (280-320nm), UV-A (320-400 nm) and visible (400 nm and above) radiation. Inparticular, Bache et al. compositions can be cured using radiationhaving a wavelength of 320 nm or more. When fully cured (regardless ofthe type of radiation used), the Bach et al. compositions exhibithardnesses and impact resistances at least comparable to conventionalcoatings.

In the invention described here, the luminescing particles (or energymodulation agents) described above are added to these Bach et al.compositions, optionally including in one embodiment various colorpigments. Due to the fact that the exterior energy source penetratesthroughout the entirety of the Bach et al. compositions, thicker surfacecoatings can be realized. Further, the coatings can be applied tointricate surfaces having for example been prepared with recesses orprotrusions. Curing with the recesses and around the protrusions withoutbeing limited by conventional UV shading will likely provide enhancedadherence of the surface coating to the work piece.

System Implementation

In one embodiment of the invention, there is provided a first system forproducing a change in a medium.

In one embodiment, the energy modulation agent converts the appliedinitiation energy and produces light at an energy different from theapplied initiation energy. In one embodiment, the applied initiationenergy source is an external initiation energy source. In oneembodiment, the applied initiation energy source is a source that is atleast partially in a container holding the medium.

The medium in one embodiment is substantially transparent to theinitiation energy. For example, if the medium is a liquid or fluid foodproduct such as orange juice which has a substantial amount of suspendedsolids, then UV light for example as described above and even visiblelight will be substantially absorbed and/or scattered by the orangejuice medium. Furthermore, microwave energy will likewise be absorbed bythis medium. However, an initiation energy source such as an X-raysource will essentially transmit entirely through for example an orangejuice medium. The effect is the medium can now be totally illuminatedwith the external initiation energy source.

Other sources and tuned to specific wavelengths may also be used as theinitiation energy source. These sources would take advantage of an“optical window” in the medium where for example a particular wavelengthof light would not be absorbed. Water selectively scatters and absorbscertain wavelengths of visible light. The long wavelengths of the lightspectrum—red, yellow, and orange—can penetrate to approximately 15, 30,and 50 meters (49, 98, and 164 feet), respectively, while the shortwavelengths of the light spectrum—violet, blue and green—can penetratefurther. Thus, for many aqueous based systems, non-high energy X-raysources may not be needed. In those situations, energy modulation agentswould be added whose interaction with the incident light would producefor example photoactivation of catalysts in the aqueous medium.

Accordingly, depending on the medium and the energy modulation agent andthe activatable agent, the initiation energy source can include at leastone of an X-ray source, a gamma ray source, an electron beam source, anUV radiation source, a visible and infrared source, a microwave source,or a radio wave source. The initiation energy source can then be anenergy source emitting one of electromagnetic energy, acoustic energy,or thermal energy. The initiation energy source can then be an energysource emitting a wavelength whose depth of penetration penetratesthroughout the medium. The medium to be effected can be a medium to befermented, sterilized, or cold pasteurized. The medium to be effectedcan include bacteria, viruses, yeasts, and fungi.

The activatable agents can be photoactivatable agents such as thephotocages (described elsewhere) such that upon exposure to theinitiation energy source, the photocage disassociates rendering anactive agent available. The activatable agents can include agents suchas psoralens, pyrene cholesteryloleate, acridine, porphyrin,fluorescein, rhodamine, 16-diazorcortisone, ethidium, transition metalcomplexes of bleomycin, transition metal complexes of deglycobleomycinorganoplatinum complexes, alloxazines, vitamin Ks, vitamin L, vitaminmetabolites, vitamin precursors, naphthoquinones, naphthalenes,naphthols and derivatives thereof having planar molecular conformations,porphorinporphyrins, dyes and phenothiazine derivatives, coumarins,quinolones, quinones, and anthroquinones. The activatable agents caninclude photocatalysts such as TiO₂, ZnO, CdS, CdSe, SnO₂, SrTiO₃, WO₃,Fe₂O₃, and Ta₂O₅ particles.

The first system can include a mechanism configured to provide in themedium at least one energy modulation agent which converts theinitiation energy to an activation energy for activation of theactivatable agent(s). The energy modulation agent(s) can be a photonemitter such a phosphorescent compounds, chemiluminescent compounds, andbioluminescent compounds. The energy modulation agent(s) can be upconversion or down conversion agents. The energy modulation agent(s) canbe luminescent particles which emit light upon exposure to saidinitiation energy. The energy modulation agent(s) can be nanotubes,nanoparticles, chemilumiscent particles, and bioluminescent particles,and mixtures thereof. The luminescent particles can be chemiluminescentparticles which show enhanced chemiluminescence upon exposure tomicrowaves.

Depending on the initiation energy source, the system can include acontainer for the medium that is permeable to the applied initiationenergy. For example, for an X-ray source, the container can be made ofaluminum, quartz, glass, or plastic. For a microwave source, thecontainer can be made of quartz, glass, or plastic. Furthermore, thecontainer can be a container which receives and transmits the initiationenergy to fluid products to pasteurize the fluid products, or can be acontainer which receives and transmits the initiation energy to fluidproducts to remediate contaminants in the fluid products.

In another embodiment of the invention, there is provided a secondsystem for curing a radiation-curable medium. The second system includesa mechanism configured to supply an uncured radiation-curable mediumincluding at least one activatable agent which produces a change in theradiation-curable medium when activated, and further includes an appliedinitiation energy source configured to apply initiation energy to acomposition including the uncured radiation-curable medium and theenergy modulation agent. The energy modulation agent as described aboveabsorbs the initiation energy and converts the initiation energy to anactivation energy capable of curing the uncured medium (i.e., promotingpolymerization of polymers in the uncured medium). In another example,activation of the energy modulation agent produces a light whichactivates the at least one photoactivatable agent to polymerize polymersin the medium.

The second system has attributes similar to the first system describedabove and can further permit the at least one activatable agent toinclude a photoinitiator such as one of benzoin, substituted benzoins,alkyl ester substituted benzoins, Michler's ketone,dialkoxyacetophenones, diethoxyacetophenone, benzophenone, substitutedbenzophenones, acetophenone, substituted acetophenones, xanthone,substituted xanthones, benzoin methyl ether, benzoin ethyl ether,benzoin isopropyl ether, diethoxyxanthone, chloro-thio-xanthone,azo-bisisobutyronitrile, N-methyl diethanolaminebenzophenone,camphoquinone, peroxyester initiators, non-fluorene-carboxylic acidperoxyesters and mixtures thereof.

The second system can include a container for the uncuredradiation-curable medium that is permeable to the applied initiationenergy. The container can be configured to contain the uncuredradiation-curable medium or to hold a mold of the uncuredradiation-curable medium. The container as before can be an aluminumcontainer, a quartz container, a glass container, or a plasticcontainer, depending on the applied initiation energy.

In one embodiment, an energy source (e.g., an external energy source) isconfigured to irradiate the uncured radiation-curable medium in a jointregion (or regions) adhering one region of a utensil to another regionof the utensil. In another embodiment, the energy source is configuredto irradiate the joint regions and thereby induce sterilization of thejoint regions due to the production of internal UV light inside thejoint regions. In another embodiment, the energy source is configured toirradiate a surface coating.

The radiation-curable medium in the surface coating or in the mold or inother medium can include color pigments to add color to a finished curedproduct. The radiation-curable medium in the surface coating or in themold or in another medium can include fumed silica to promote strengthand enhance distribution of the internally generated light. Theradiation-curable medium in the surface coating or in the mold or inanother medium can include a moisture cure promoter to supplement thecure.

The second system provides one mechanism for production of novelradiation-cured articles, which include a radiation-cured medium and atleast one energy modulation agent distributed throughout the medium. Theenergy modulation agent being a substance which is capable of convertingan applied energy to light capable of producing a cure for theradiation-cured medium. The article can include luminescent particlessuch as for example nanotubes, nanoparticles, chemilumiscent particles,and bioluminescent particles, and mixtures thereof. The article caninclude chemiluminescent particles. The article can include colorpigments or fumed silica.

In another embodiment of the invention, there is provided a third systemfor producing a change in a medium disposed in an artificial container.The third system includes a mechanism configured to provide to themedium 1) an activatable agent and 2) at least one energy modulationagent. The energy modulation agent converts an initiation energy to anactivation energy which then activates the at least one activatableagent. The third system further includes an applied initiation energysource configured to apply the initiation energy through the artificialcontainer to activate the at least one activatable agent in the medium.

The third system has similar attributes to the first and second systemsdescribed above, and further includes encapsulated structures includingthe energy modulation agent. The encapsulated structures can includenanoparticles of the energy modulation agent encapsulated with apassivation layer or can include sealed quartz or glass tubes having theenergy modulation agent inside.

In another embodiment of the invention, there is provided a fourthsystem for producing a photo-stimulated change in a medium disposed inan artificial container. The fourth system includes a mechanismconfigured to provide in the medium at least one energy modulationagent. The energy modulation agent converts an initiation energy to anactivation energy which then produces the photo-stimulated change. Thefourth system further includes an initiation energy source configured toapply the initiation energy to the medium to activate the at least oneenergy modulation agent in the medium. The system can includeencapsulated structures including therein the energy modulation agent.The encapsulated structures can include nanoparticles of the energymodulation agent encapsulated with a passivation layer.

The fourth system can include a container which receives and transmitsthe initiation energy to products within the medium. The products caninclude plastics, where the activation energy alters the surfacestructure of the plastics. The products can include polylactic acid(PLA) plastics and polyhydroxyalkanoates (PHA) plastics. In thisembodiment, the activation energy can photo-graft a molecular speciesonto a surface of the plastics.

Sterilization Methods and System Components

Optical techniques have been often used in sterilization procedures torender unwanted or harmful waterborne microorganisms incapable ofreproducing using ultraviolet light (specifically the spectral area ofUV-C, 200 to 280 nm range). Ultraviolet light in the UV-C is consideredthe most lethal range as a germicidal disinfectant (capable of alteringa living microorganism's DNA, and keeping the microorganism fromreproducing). UV-C, with 264 nanometers being the peak germicidalwavelength, is known as the germicidal spectrum. Although the UV-Cmethod is simple and effective, it is not particularly effective insamples (gas, liquids, particulates) enclosed on containers which do nottransmit UV light. The present invention provides techniques and systemsthat can use externally applied radiation such as X-ray forsterilization. While illustrated below with respect to X-rayirradiation, and as discussed above, other suitable forms of energycould be used provided the containers and medium to be sterilized wassufficiently transparent for the medium to be thoroughly irradiated.Examples of alternative sources and materials for upconvertingluminescence to higher energies have been discussed above.

These systems are applicable in a number of the applications discussedabove and as well as in other sterilization areas. The systems couldthus be used in the waste water detoxification, blood sterilization,cold pasteurization, and photodeactivation commercial applicationsdiscussed in the sections above. These systems (like FIGS. 3B-3D) showthe use of artificial containers in which the medium to be treated isdisposed.

Having generally described this invention, a further understanding canbe obtained by reference to certain specific examples which are providedherein for purposes of illustration only and are not intended to belimiting unless otherwise specified.

Numerous modifications and variations of the invention are possible inlight of the above teachings. It is therefore to be understood thatwithin the scope of the appended claims, the invention may be practicedotherwise than as specifically described herein.

The invention claimed is:
 1. A method for treating a condition, disorderor disease comprising: providing an activation energy sufficient tocause a predetermined change in a subject, wherein the activation energyis generated in-situ in said subject by application of an initiationenergy which is converted in-situ to said activation energy, wherein theinitiation energy is at least one member selected from the groupconsisting of x-rays, gamma rays, and electron beams; and prior to orsimultaneously with providing the activation energy, administering tothe subject at least one activatable pharmaceutical agent that isactivatable in situ by the activation energy, wherein upon activation,the activated at least one activatable pharmaceutical agent effects thepredetermined change in the subject, wherein said predetermined changeat least one of 1) causes a change in organism activity or 2) treats thecondition, disorder, or disease.
 2. The method of claim 1, furthercomprising, prior to said applying of the initiation energy,administering to the subject at least one energy modulation agent thatassists in converting the initiation energy to the activation energy. 3.The method of claim 2, wherein the at least one energy modulation agentcomprises one or more selected from a biocompatible fluorescing metalnanoparticle, fluorescing dye molecule, gold nanoparticle, a watersoluble quantum dot encapsulated by polyamidoamine dendrimers, aluciferase, a biocompatible phosphorescent molecule, a combinedelectromagnetic energy harvester molecule, and a lanthanide chelatecapable of intense luminescence.
 4. The method of claim 2, wherein theenergy modulation agent comprises a fluorophore.
 5. The method of claim2 wherein the at least one energy modulation agent comprises a singleenergy modulation agent, and is coupled to said at least one activatablepharmaceutical agent.
 6. The method of claim 1, wherein the initiationenergy source is selected from the group consisting of x-rays andelectron beams.
 7. The method of claim 1, wherein the initiation energyproduces energy states for energy transfer to the at least oneactivatable pharmaceutical agent.
 8. The method of claim 1, wherein theinitiation energy is directed at a solid tumor.
 9. The method of claim1, wherein the initiation energy directly or indirectly activates the atleast one activatable pharmaceutical agent.
 10. The method of claim 1,wherein the initiation energy directly or indirectly activates apsoralen.
 11. The method of claim 10, wherein the initiation energydirectly or indirectly activates 8-MOP or AMT.
 12. The method of claim11, wherein the initiation energy damages DNA in target cells.
 13. Themethod of claim 1, wherein the initiation energy comprises ionizingradiation which interacts to produce reactive groups at a target site.14. The method of claim 13, wherein the reactive groups comprise atleast a member selected from the group consisting of ions, singletoxygen, free radicals, and hydroxides.
 15. The method of claim 1,wherein the predetermined change is a predetermined cellular change. 16.The method of claim 15, wherein said condition, disorder or disease is acell proliferation disorder and said predetermined cellular changetreats the cell proliferation disorder by causing an increase ordecrease in cell proliferation rate of a target cell.
 17. The method ofclaim 1, wherein the at least one activatable pharmaceutical agent isactivated by one or more sequential single photon absorption events. 18.The method of claim 1, wherein the at least one activatablepharmaceutical agent comprises an active agent contained within aphotocage, wherein upon exposure to said initiation energy, thephotocage disassociates from the active agent, rendering the activeagent available.
 19. The method of claim 1, wherein the at least oneactivatable pharmaceutical agent comprises an active agent containedwithin a photocage, and said activation energy is UV-A or visibleenergy, wherein upon exposure to said UV-A or visible energy, thephotocage disassociates from the active agent, rendering the activeagent available.
 20. The method of claim 1, the activation energy iscapable of penetrating human tissue up to about 4 mm.
 21. The method ofclaim 1, wherein the at least one activated pharmaceutical agent causesan auto-vaccine effect in the subject that reacts with a target cell.22. The method of claim 1, wherein the predetermined cellular change isapoptosis in a target cell.
 23. The method of claim 1, wherein thedisorder is at least one member selected from the group consisting ofcancer, bacterial infection, viral infection, immune rejection response,autoimmune disorders, and aplastic conditions.
 24. The method of claim1, wherein the at least one activatable pharmaceutical agent is aphotoactivatable agent.
 25. The method of claim 1, wherein the at leastone activatable pharmaceutical agent is selected from psoralens, pyrenecholesteryloleate, acridine, porphyrin, fluorescein, rhodamine,16-diazorcortisone, ethidium, transition metal complexes of bleomycin,transition metal complexes of deglycobleomycin organoplatinum complexes,alloxazines, vitamin Ks, vitamin L, vitamin metabolites, vitaminprecursors, naphthoquinones, naphthalenes, naphthols and derivativesthereof having planar molecular conformations, porphyrins, dyes andphenothiazine derivatives, coumarins, quinolones, quinones, andanthroquinones.
 26. The method of claim 1, wherein the at least oneactivatable pharmaceutical agent is a psoralen, a coumarin, or aderivative thereof.
 27. The method of claim 26, wherein the at least oneactivatable pharmaceutical agent is 8-MOP or AMT.
 28. The method ofclaim 1, wherein the at least one activatable pharmaceutical agent isone selected from 7,8-dimethyl-10-ribityl, isoalloxazine,7,8,10-trimethylisoalloxazine, 7,8-dimethylalloxazine,isoalloxazine-adenine dinucleotide, alloxazine mononucleotide, aluminum(III) phthalocyanine tetrasulonate, hematoporphyrin, and phthadocyanine.29. The method of claim 1, wherein the at least one activatablepharmaceutical agent is coupled to a carrier that is capable of bindingto a receptor site.
 30. The method of claim 29, wherein the carrier isone selected from insulin, interleukin, thymopoietin or transferrin. 31.The method of claim 30, wherein the at least one activatablepharmaceutical agent is coupled to the carrier by a covalent bond. 32.The method of claim 30, wherein the at least one activatablepharmaceutical agent is coupled to the carrier by a non-covalent bond.33. The method of claim 30, wherein the receptor site is one selectedfrom nucleic acids of nucleated cells, antigenic sites on nucleatedcells, or epitopes.
 34. The method of claim 1, wherein the at least oneactivatable pharmaceutical agent has affinity for a target cell.
 35. Themethod of claim 1, wherein the at least one activatable pharmaceuticalagent is capable of being preferentially absorbed by a target cell. 36.The method of claim 1, wherein the at least one activatablepharmaceutical agent is a DNA intercalator or a halogenated derivativethereof.
 37. The method of claim 1, further comprising providing to thesubject at least one additive having a complementary therapeutic ordiagnostic effect.
 38. The method of claim 37, wherein said additivecomprises at least one of an antioxidant, an adjuvant, or a combinationthereof.
 39. The method of claim 38, wherein said antioxidant is addedto reduce side-effects of irradiation.
 40. A method for treating acondition, disorder or disease comprising: providing an activationenergy sufficient to cause a predetermined change in a subject, whereinthe activation energy is generated in-situ in said subject byapplication of an initiation energy which is converted in-situ to saidactivation energy, wherein the initiation energy is at least one memberselected from the group consisting of x-rays, gamma rays, electronbeams; and protons, and prior to or simultaneously with providing theactivation energy, administering to the subject at least one activatablepharmaceutical agent that is activatable by the activation energy,wherein upon activation, the activated at least one activatablepharmaceutical agent effects the predetermined change in the subject,wherein said predetermined change at least one of 1) causes a change inorganism activity or 2) treats the condition, disorder, or disease. 41.A method for treating a condition, disorder or disease comprising:providing an activation energy sufficient to cause a predeterminedchange in a subject, wherein the activation energy is generated in-situin said subject by application of an initiation energy which producesmultiple energy states, at least one of the energy states providing theactivation energy, wherein the initiation energy is at least one memberselected from the group consisting of x-rays, gamma rays, and electronbeams; and prior to or simultaneously with providing the activationenergy, administering to the subject at least one activatablepharmaceutical agent that is activatable by the activation energy,wherein upon activation, the activated at least one activatablepharmaceutical agent effects the predetermined change in the subject,wherein said predetermined change at least one of 1) causes a change inorganism activity 2) treats the condition, disorder, or disease.